(4-oxo-2-pyrimidinyl)thioalkyl compounds useful as AICARFT inhibitors

ABSTRACT

Compounds of the formula:                    
     (where R 1 , R 2  and R 3  are defined in the specification) are inhibitors of AICARFT. These compounds, as well as their pharmaceutically acceptable salts, solvents, prodrugs, and pharmaceutically active metabolites, are useful in pharmaceutical compositions for treating diseases such as cancer.

This application is a national phase filing under 35 U.S.C. § 371 ofInternational Patent Application No. PCT/US99/20331, filed Sep. 3, 1999,and a continuation-in-part of U.S. Provisional Patent Application No.60/099,259, filed Sep. 4, 1998, the disclosures of each of which areincorporated herein by reference.

FIELD AND INDUSTRIAL APPLICABILITY OF THE INVENTION

The present invention relates to compounds that inhibit the enzymeaminoimidazole carboxamide ribonucleotide formyl transferase (AICARFT),to pharmaceutical compositions containing these compounds, and to theiruse to inhibit AICARFT. The-invention also relates to the preparation ofthese compounds, and to intermediates for preparing these compounds.

BACKGROUND OF THE INVENTION

The large class of antiproliferative agents includes antimetabolitecompounds. A particular subclass of antimetabolites known as antifolatesor antifoles are antagonists of the vitamin folic acid.

Aminoimidazole carboxamide ribonucleotide formyl transferase (AICARFT)is a folate-dependent enzyme in the de novo purine biosynthesis pathway.This pathway is critical to cell division and proliferation. Shuttingdown this pathway is known to have an antiproliferative, in particular,an antitumor effect.

There is a need for compounds that inhibit the enzyme AICARFT, havingantitumor, antiinflammatory, antipsoriatic, and/or immunosuppressiveactivity.

SUMMARY OF THE INVENTION

An object of the invention is to provide small-molecule compounds thatinhibit AICARFT. Another object of the invention is to provideantitumor, antiinflammatory, antipsoriatic, or immunosuppressive agentsuseful in pharmaceutical treatments.

Surprisingly, such objects have been achieved by the compounds offormula I:

wherein:

R¹ is H or CN;

R² is phenyl or thienyl, each of which may be optionally substitutedwith phenyl, phenoxy, thienyl, tetrazolyl, or 4-morpholinyl; and

R³ is phenyl substituted with —SO₂NR⁵R⁶ or —NR⁵SO₂R⁶ and optionallyfurther substituted by one or more of the suitable substituents definedbelow; wherein R⁵ is H or lower alkyl, R⁶ is lower alkyl, substituted orunsubstituted heteroarylalkyl, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl, wherein the aryl and heteroarylmoieties may be substituted by one or more of the suitable substituentsdefined below; or

 wherein n is an integer of from 1 to 4, R⁴ is OH, lower alkoxy, or aglutamic-acid or glutamate-ester moiety linked through the aminefunctional group.

The invention also relates to pharmaceutically acceptable salts,solvates, prodrugs, and active metabolites of compounds of the formulaI.

In preferred embodiments, R² is selected from

and/or

R³ may be phenyl substituted with —SO₂NR⁵R⁶ or —NR⁵SO₂R⁶ and optionallyfurther substituted by lower alkyl, lower alkoxy, or halogen. Forexample, R³ may be:

each of which may be optionally substituted with one additionalsubstituent selected from methyl, methoxy and chloro.

Perferred compounds of the invention are:

and pharmaceutically acceptable salts, solvates, prodrugs, and activemetabolites of such compounds.

The invention further relates to use of the compounds of formula I, andespecially the above-described preferred compounds, as inhibitors of theenzyme AICARFT.

The invention also relates to methods of synthesizing the compounds ofthe invention, comprising carrying out the following general alkylationreaction:

where R¹, R², and R³ are as defined above.

Other features, objects, and advantages of the invention will becomeapparent from the detailed description of the invention provided below.

DETAILED DESCRIPTION AND PREFERRED EMBODIMENTS

In accordance with a convention used in the art, is used in structuralformulae herein to depict the bond that is the point of attachment ofthe moiety or substituent to the core or backbone structure.

Where chiral carbons are included in chemical structures, unless aparticular orientation is depicted, both stereoisomeric forms areintended to be encompassed. Further, the inventive compounds may existas single stereoisomers, racemates, and/or mixtures of enantiomersand/or diastereomers. All such single stereoisomers, racemates, andmixtures thereof are intended to be within the broad scope of thepresent invention. Preferably, however, the inventive compounds are usedin optically pure form.

As generally understood by those skilled in the art, an optically purecompound is one that is essentially enantiomerically pure. Preferably,an optically pure compound of the invention contains at least 90% of asingle isomer (80% enantiomeric excess), more preferably at least 95%(90% e.e.), even more preferably at least 97.5% (95% e.e.), and mostpreferably at least 99% (98% e.e.).

The compounds illustrated by the chemical formulae referred to hereinmay exhibit the phenomenon of tautomerism. Although the structuralformulae depict one of the possible tautomeric forms, it should beunderstood that the invention nonetheless encompasses all tautomericforms.

As used herein, the term “alkyl group” is intended to mean a straight-or branched-chain monovalent radical of saturated and/or unsaturatedcarbon atoms and hydrogen atoms, such as methyl (Me), ethyl (Et),propyl, isopropyl, butyl, isobutyl, t-butyl, ethenyl, pentenyl, butenyl,propenyl, ethynyl, butynyl, propynyl, pentynyl, hexynyl, and the like,which may be unsubstituted (i.e., containing only carbon and hydrogen)or substituted by one or more suitable substituents as defined below(e.g., one or more halogens, such as F, Cl, Br, or I, with F and Clbeing preferred). A “lower alkyl group” is intended to mean an alkylgroup having from 1 to 4 carbon atoms in its chain.

An “alkoxy group” is intended to mean the radical —OR_(a), where R_(a)is an alkyl group. Exemplary alkoxy groups include methoxy, ethoxy, andpropoxy. “Lower alkoxy” refers to alkoxy groups wherein the alkylportion has 1 to 4 carbon atoms.

An “aryl” group is intended to mean a cyclic aromatic hydrocarbon group,such as phenyl or naphthyl, which may be unsubstituted or substituted byone or more of the suitable substituents defined below (e.g., with oneor more halogen, lower alkyl, and/or lower alkoxy group). Preferably,aryl is a substituted or unsubstituted phenyl group.

A “heteroaryl” group is intended to mean a cyclic aromatic groupcontaining at least one heteroatom selected from oxygen, sulfur andnitrogen, wherein any position of the heteroaryl group may beunsubstituted or substituted by a suitable substituent, as defined below(e.g., with a halogen, lower alkyl, or lower alkoxy group). Preferablythe heteroaryl group is a nitrogen-containing cyclic aromatic group.Exemplary nitrogen-containing heteroaryl groups include quinolyl,isoquinolyl, indolyl, pyridyl, pyrrolyl, pyrazinyl, and thiazolyl. A“heteroarylalkyl” group comprises an alkylenyl moiety bonded to asubstituted or unsubstituted heteroaryl group, e.g., —(CH₂)_(x)-indolyl,wherein x may be an integer of from 1 to 4 and any position of theheteroaryl group may be unsubstituted or substituted by a suitablesubstituent, as defined below. Preferable heteroaryl groups includesubstituted or unsubstituted isoquinolyl or indolyl groups.

The term “substituent” or “suitable substituent” is intended to mean anysuitable substituent that may be recognized or selected, such as throughroutine testing, by those skilled in the art. Illustrative examples ofsuitable substituents include hydroxy groups, halogens, oxo groups,alkyl groups, acyl groups, sulfonyl groups, mercapto groups, alkylthiogroups, alkoxy groups, cycloalkyl groups, heterocycloalkyl groups, arylgroups, heteroaryl groups, carboxy groups, amino groups, alkylaminogroups, dialkylamino groups, carbamoyl groups, aryloxy groups,heteroaryloxy groups, arylthio groups, heteroarylthio groups, and thelike.

The term “optionally substituted” is intended to expressly indicate thatthe specified group is unsubstituted or substituted with one or moresuitable substituents, unless the optional substituents are expresslyspecified, in which case the term indicates that the group isunsubstituted or substituted with the specified substituents. As definedabove, various groups may be unsubstituted or substituted (i.e., theyare optionally substituted) unless indicated otherwise herein (e.g., byindicating that the specified group is unsubstituted).

A “prodrug” is intended to mean a compound that is converted underphysiological conditions or by solvolysis or metabolically to aspecified compound that is pharmaceutically active.

A “solvate” is intended to mean a pharmaceutically acceptable solvateform of a specified compound that retains the biological effectivenessof such compound. Examples of solvates include compounds of theinvention in combination with water, isopropanol, ethanol, methanol,DMSO, ethyl acetate, acetic acid, or ethanolamine.

A “pharmaceutically active metabolite” is intended to mean apharmacologically active product produced through metabolism in the bodyof a specified compound.

A “pharmaceutically acceptable salt” is intended to mean a salt thatretains the biological effectiveness of the free acids and bases of thespecified compound and that is not biologically or otherwiseundesirable. Examples of pharmaceutically acceptable salts includesulfates, pyrosulfates, bisulfates, sulfites, bisulfites, phosphates,monohydrogenphosphates, dihydrogenphosphates, metaphosphates,pyrophosphates, chlorides, bromides, iodides, acetates, propionates,decanoates, caprylates, acrylates, formnates, isobutyrates, caproates,heptanoates, propiolates, oxalates, malonates, succinates, suberates,sebacates, fumarates, maleates, butyne-1,4-dioates, hexyne-1,6-dioates,benzoates, chlorobenzoates, methylbenzoates, dinitrobenzoates,hydroxybenzoates, methoxybenzoates, phthalates, sulfonates,xylenesulfonates, phenylacetates, phenylpropionates, phenylbutyrates,citrates, lactates, γ-hydroxybutyrates, glycollates, tartrates,methane-sulfonates, propanesulfonates, naphthalene-1-sulfonates,naphthalene-2-sulfonates, mandelates, and mesylates.

If a compound is a base, a desired salt may be prepared by any suitablemethod known in the art, including treatment of the free base with aninorganic acid, such as hydrochloric acid; hydrobromic acid; sulfuricacid; nitric acid; phosphoric acid; and the like; or with an organicacid, such as acetic acid; maleic acid; succinic acid; mandelic acid;fumaric acid; malonic acid; pyruvic acid; oxalic acid; glycolic acid;salicylic acid; pyranosidyl acid, such as glucuronic acid orgalacturonic acid; alpha-hydroxy acid, such as citric acid or tartaricacid; amino acid, such as aspartic acid or glutamic acid; aromatic acid,such as benzoic acid or cinnamic acid; sulfonic acid, such asp-toluenesulfonic acid or ethanesulfonic acid; or the like.

If a compound is an acid, a desired salt may be prepared by any suitablemethod known in the art, including treatment of the free acid with aninorganic or organic base, such as an amine (primary, secondary, ortertiary); an alkali metal or alkaline earth metal hydroxide; or thelike. Illustrative examples of suitable salts include organic saltsderived from amino acids such as glycine and arginine; ammonia; primary,secondary, and tertiary amines; and cyclic amines, such as piperidine,morpholine, and piperazine; as well as inorganic salts derived fromsodium, calcium, potassium, magnesium, manganese, iron, copper, zinc,aluminum, and lithium.

In the case of compounds, salts, or solvates that are solids, it isunderstood by those skilled in the art that the inventive compounds,salts, and solvates may exist in different crystal forms, all of whichare intended to be within the scope of the present invention andspecified formulae.

The present invention is also directed to a method of inhibiting AICARFTactivity, comprising contacting the enzyme with an effective amount of acompound of formula I, or a pharmaceutically acceptable salt, prodrug,pharmaceutically active metabolite, or solvate thereof. For example,AICARFT activity may be inhibited in mammalian tissue by administering acompound of formula I or a pharmaceutically acceptable salt, prodrug,pharmaceutically active metabolite, or solvate thereof. The compounds offormula I may also be used to inhibit the growth or proliferation ofviruses and/or cells of higher organisms or microorganisms such asyeast, bacteria and fungi. The invention is also directed to methods oftreating cancer by administering to a patient in need of such treatmentan effective amount of a compound of the formula I, or apharmaceutically acceptable salt, prodrug, pharmaceutically activemetabolite, or solvate thereof.

“Treating” or “treatment” is intended to mean at least the mitigation ofa disease condition in a mammal, such as a human, that is mediated bythe inhibition of the activity of the target (e.g., reduction in tumorgrowth), and includes: (a) prophylactic treatment in a mammal,particularly when the mammal is found to be predisposed to having thedisease condition but not yet diagnosed as having it; (b) inhibiting thedisease condition; and/or (c) alleviating, in whole or in part, thedisease condition.

The potency of the inventive compounds as inhibitors of AICARFT activitymay be measured by any of the suitable methods known to those skilled inthe art, including in vivo and in vitro assays. An example of a suitableassay for activity measurements is the AICARFT enzyme assay describedherein.

Administration of the compounds of the formula I and theirpharmaceutically acceptable prodrugs, salts, active metabolites, andsolvates may be performed according to any of the accepted modes ofadministration available to those skilled in the art. Illustrativeexamples of suitable modes of administration include oral, nasal,parenteral, topical, transdermal, and rectal.

An inventive compound of formula I or a pharmaceutically acceptablesalt, prodrug, active metabolite, or solvate thereof may be administeredas a pharmaceutical composition in any pharmaceutical form recognizableto the skilled artisan as being suitable. Suitable pharmaceutical formsinclude solid, semisolid, liquid, or lyophilized formulations, such astablets, powders, capsules, suppositories, suspensions, liposomes, andaerosols. Pharmaceutical compositions of the invention may also includesuitable excipients, diluents, vehicles, and carriers, as well as otherpharmaceutically active agents, depending upon the intended use.

Acceptable methods of preparing suitable pharmaceutical forms of thepharmaceutical compositions are known or may be routinely determined bythose skilled in the art. For example, pharmaceutical preparations maybe prepared following conventional techniques of the pharmaceuticalchemist involving steps such as mixing, granulating, and compressingwhen necessary for tablet forms, or mixing, filling, and dissolving theingredients as appropriate, to give the desired products for oral,parenteral, topical, intravaginal, intranasal, intrabronchial,intraocular, intraaural, and/or rectal administration.

Solid or liquid pharmaceutically acceptable carriers, diluents,vehicles, or excipients may be employed in the pharmaceuticalcompositions. Illustrative solid carriers include starch, lactose,calcium sulfate dihydrate, terra alba, sucrose, talc, gelatin, pectin,acacia, magnesium stearate, and stearic acid. Illustrative liquidcarriers include syrup, peanut oil, olive oil, saline solution, andwater. The carrier or diluent may include a suitable prolonged-releasematerial, such as glyceryl monostearate or glyceryl distearate, alone orwith a wax. When a liquid carrier is used, the preparation may be in theform of a syrup, elixir, emulsion, soft gelatin capsule, sterileinjectable liquid (e.g., solution), or a nonaqueous or aqueous liquidsuspension.

A dose of the pharmaceutical composition contains at least atherapeutically effective amount of the active compound or agent (i.e.,a compound of formula I or a pharmaceutically acceptable salt, prodrug,active metabolite, or solvate thereof), and preferably is made up of oneor more pharmaceutical dosage units. The selected dose may beadministered to a mammal, for example, a human patient, in need oftreatment mediated by inhibition of AICARFT activity, by any known orsuitable method of administering the dose, including topically, forexample, as an ointment or cream; orally; rectally, for example, as asuppository; parenterally by injection; or continuously by intravaginal,intranasal, intrabronchial, intraaural, or intraocular infusion.

An “effective amount” is intended to mean a therapeutic amount of aninventive compound that, when administered to a mammal in need oftreatment, is sufficient to effect treatment for disease conditionsalleviated by the inhibition of the target (i.e., AICARFT) activity,such as tumor growth. The amount of a given compound of the inventionthat will be therapeutically effective will vary depending upon factorssuch as the particular compound, the disease condition and the severitythereof, the identity of the mammal in need of treatment, which amountmay be routinely determined by artisans.

An exemplary dose of a compound of the invention for a vertebrate hostcomprises an amount up to one gram of active compound per kilogram ofthe host, preferably one-half of a gram, more preferably 100 milligrams,and most preferably, about 50 milligrams or less, per kilogram of thehost weight.

The pharmaceutical compositions of the invention may further compriseone or more other pharmaceutically active compounds. For example, foranticancer compositions, one of the following antitumor agents may beincluded: mitotic inhibitors (e.g., vinblastine); alkylating agents;dihydrofolate reductase inhibitors or thymidylate synthase inhibitors;antimetabolites (e.g., 5-fluorouracil, cytosinearabinoside);intercalating antibiotics (e.g. adriamycin, bleomycin); enyzmes (e.g.,asparaginase); topoisomerase inhibitors (e.g., etoposide); andbiological-response modifiers (e.g., interferon). The compositions ofthe invention may also comprise another enzyme inhibitor such as a GARFTinhibitor or antiproliferative agent, such as a compound described inU.S. Pat. No. 5,610,319 or U.S. Pat. No. 5,574,039, the disclosures ofwhich are herein incorporated by reference. The compositions of theinvention may also comprise one or more antibacterial, antifungal,antiparasitic, antiviral, antipsoriatic, or anticoccidial agent.Exemplary antibacterial agents include sulfonamides, such assulfamethoxazole, sulfadiazine, sulfameter, or sulfadoxine; dihydrofolicreductase inhibitors, such as trimethoprim, bromodiaprim, ortrimetrexate; penicillins; cephalosporins; and the quinolone carboxylicacids and their fused isothiazolo analogs.

The skeletal framework for the molecules of formula I may be assembledby the alkylation of the appropriate arylthiol with the correspondingalkyl bromide:

5-[(Arylthio)alkyl]thiophene-2-carboxylic acids and the5-[(arylthio)alkyl]theno-2-yl glutamatic acids are prepared by thebase-catalyzed hydrolysis of the requisite methyl esters and diethylesters, respectively.

6-Phenyl-2-thiouracil is a commercially available aryl thiol; the other6-aryl-4(3H)-oxopyrimidine-2-thiols are prepared by base-catalyzedcondensation of the corresponding aroylacetate with thiourea.5-Cyano-6-[5-(2-thienyl)thien-2-yl]-4(3H)-oxopyrimidine-2-thiol issynthesized from the condensation of equimolar amounts of2,2′-bithiophene-5-carboxaldehyde, thiourea, ethyl cyanoacetate, andpotassium carbonate.

The alkyl bromides are prepared from their alcohol precursors bytreatment with carbon tetrabromide and triphenyl phosphine. In general,the 5-(hydroxyalkyl)thiophene-2-carboxylates and the[5-(hydroxyalkyl)theno-2yl]glutamates are prepared by thepalladium-catalyzed coupling of the requisite hydroxyalkyne with eithermethyl 5-bromothiophene-2-carboxylate or diethylN-(5-bromotheno-2-yl)glutamate, followed by hydrogenation of the triplebond. The (hydroxymethyl)bisaryl sulfonamides are generally prepared byreduction of the corresponding methyl esters, which may be synthesizedby sulfonylation of the appropriate aniline with the requisite sulfonylchloride.

EXAMPLES

Specific examples of various preferred compounds of formula I and theirpreparation are described below. The structures of the compounds of thefollowing examples were confirmed by one or more of the following:proton magnetic resonance spectroscopy, infrared spectroscopy, elementalmicroanalysis, mass spectrometry, thin layer chromatography,melting-point determination, and boiling-point determination. Wherethere is any discrepancy between the given structural formula shown fora compound and its chemical name provided, the structural formula isintended to apply.

Proton magnetic resonance (¹H NMR) spectra were determined using eithera Bruker DPX 300 or a General Electric QE-300 spectrometer operating ata field strength of 300 megahertz (MHz). Chemical shifts are reported inparts per million (ppm, δ) downfield from an internal tetramethylsilanestandard. Alternatively, ¹H NMR spectra were referenced to residualprotic solvent signals as follows: CHCl₃=7.26 ppm; DMSO=2.49 ppm. Peakmultiplicities are designated as follows: s=singlet; d=doublet;dd=doublet of doublets; ddd=doublet of doublet of doublets; t=triplet;tt=triplet of triplets; q=quartet; br=broad resonance; and m=multiplet.Coupling constants are given in Hertz (Hz). Infrared absorption (IR)spectra were obtained using a Perkin-Elmer 1600 series-FTIRspectrometer. Elemental microanalyses were performed (by AtlanticMicrolab Inc., Norcross, Ga.) and gave results for the elements statedwithin ±0.4% of the theoretical values. Flash column chromatography wasperformed using Silica Gel 60 (Merck Art 9385). Analytical thin layerchromatography (TLC) was performed using precoated sheets of Silica 60F₂₅₄ (Merck Art 5719). Melting points (mp) were determined on a Mel-Tempapparatus and are uncorrected. All reactions were performed inseptum-sealed flasks under a slight positive pressure of argon, unlessotherwise noted. All commercial reagents were used as received fromtheir respective suppliers.

The following abbreviations are used herein: RT denotes roomtemperature; Et₂O refers to diethyl ether: DMF refers toN,N-dimethylformamide; and DMSO refers to dimethylsulfoxide. Otherabbreviations include: CH₃OH (methanol), EtOH (ethanol), EtOAc (ethylacetate), DME (ethylene glycol dimethyl ether), Ac (acetyl), Me(methyl), Ph (phenyl), DIEA (N,N-diisopropylethylamine), HOBt(1-hydroxybenzotriazole hydrate), EDC(1-(3-dimethylaminopropyl)-3-ethylcarbarbodiimide hydrochloride), DCC(dicyclohexylcarbodi-imide), and DMAP (4-dimethylaminopyridine).

Unless otherwise indicated, all percentages and ppm are in weight, andall temperatures are in degrees Celsius.

Example 1

(a) Methyl 5-(5-hydroxy-1-pentynyl)thiophene-2-carboxylate. To a stirredsolution of methyl 5-bromothiophene-2-carboxylate (11.05 g, 50 mmol) indiethylamine (150 mL) under an argon atmosphere were addedbis(triphenylphosphine)palladium dichloride (175 mg, 0.25 mmol), cuprousiodide (95 mg, 0.5 mmol) and 4-pentyn-1-ol (5.8 mL, 5.24 g, 62 mmol).The resulting mixture was stirred for 18 hours at ambient temperature.After removal of the solvent by concentration in vacuo, the residueobtained was diluted with water (400 mL) and extracted with EtOAc (3×150mL). The organic extracts were combined, dried over MgSO₄, andconcentrated in vacuo, to give a brown gum, which was purified by flashchromatography. Elution with hexane:EtOAc (2:1) provided the product asan orange oil (9.89 g, 88% yield). ¹H NMR (CDCl₃) δ: 7.62 (1H, d, J=3.9Hz), 7.06 (1H, d, J=3.9 Hz), 3.87 (3H, s), 3.80 (2H, t, J=6.1 Hz), 2.58(2H, t, J=7.0 Hz), 1.87 (2H, tt, J=6.1, 7.0 Hz). Anal. (C₁₁H₁₂O₃S) C, H,S.

(b) Methyl 5-(5-hydroxypentyl)thiophene-2-carboxylate. A suspension ofmethyl 5-(5-hydroxy-1-pentynyl)thiophene-2-carboxylate (9.25 g, 41.2mmol) and 5% Pd on carbon (1.39 g, 15% wt. equiv.) in EtOAc (180 mL) wasshaken under 50 psi of H₂ for 20 hours. The crude reaction mixture wasfiltered through a pad of Celite, and the filtrate was concentrated invacuo, to provide the product as a yellow oil (9.33 g, 99% yield). ¹HNMR (CDCl₃) δ: 7.63 (1H, d, J=3.7 Hz), 6.78 (1H, d, J=3.7 Hz), 3.86 (3H,s), 3.65 (2H, t, J=6.5 Hz), 2.85 (2H, t, J=7.5 Hz), 1.78-1.68 (2H, m)1.65-1.56 (2H, m), 1.49-1.39 (2H, m). Anal. (C₁₁H₁₆O₃S) C, H, S.

(c) Methyl 5-(5-bromopentyl)thiophene-2-carboxylate. A solution oftriphenylphosphine (14.43 g, 55 mmol) in 100 mL of CH₂C₂ was addeddropwise to a stirred solution of methyl5-(5-hydroxypentyl)thiophene-2-carboxylate (10.44 g, 45.7 mmol) and CBr₄(18.24 g, 55 mmol) in CH₂Cl₂ at about 0° C. over a 15-minute interval.The resulting solution was stirred at 0° C. for about 30 minutes, thenovernight at room temperature. The crude reaction mixture wassubsequently concentrated in vacuo, and the residue obtained waspurified by flash chromatography. Elution with hexane:EtOAc (95:5)provided the product as a yellow oil (12.36 g, 93% yield). ¹H NMR(CDCl₃) δ: 7.63 (1H, d, J=3.7 Hz), 6.79 (1H, d, J=3.7 Hz), 3.86 (3H, s),3.40 (2H, t, J=6.6 Hz), 2.85 (2H, t, J=7.5 Hz), 1.94-1.84 (2H, m),1.78-1.67 (2H, m), 1.56-1.47 (2H, m). Anal. (C₁₁H₁₅O₂S, Br) C, H, S, Br.

(d) Methyl5-(5-[(6phenyl-4(3H)-oxopyrimidin-2-yl)thio]pentyl)thiophene-2-carboxylate.To a stirred solution of methyl 5-(5-bromopentyl)thiophene-2-carboxylate(437 mg, 1.5 mmol) in 15 mL of DMF were added 6-phenyl-2-thiouracil (306mg, 1.5 mmol) and diisopropylethylamine (270 μl, 200 mg, 1.55 mmol).This mixture was heated at about 90° C. under an argon atmosphere forabout one hour. After cooling to room temperature, the mixture waspoured into water (100 mL). The resulting precipitate was collected byfiltration, and then washed with water (2×20 mL) and with ether (2×20mL), to provide the product as a white solid (474 mg, 76%). ¹H NMR(DMSO-d₆) δ: 12.73 (1H, br), 8.05 -8.01 (2H, m), 7.61 (1H, d, J=3.7 Hz),7.49 -7.41 (3H, m), 6.92 (1H, d, J=3.7 Hz), 6.65 (1H, s), 3.77 (3H, s),3.23 (2H, t, J=7.0 Hz), 2.83 (2H, t, J=7.4 Hz), 1.79 -1.62 (4H, m),1.50-1.41 (2H, m). Anal. (C₂₁H₂₂N₂O₃S₂) C, H, N, S.

(e)5-(5-[(6-Phenyl-4(3H)-oxopyrimidin-2-yl)thio]pentyl)thiophene2-carboxylicacid (1). A suspension of methyl5-(5-[(6-phenyl-4(3H)-oxopyrimidin-2-yl)thio]pentyl)thiophene-2-carboxylate(404 mg, 1 mmol) in 20 mL of 1N NaOH was stirred overnight at ambienttemperature, and then filtered. The filtrate was acidified to pH 4.0 byaddition of 6N HCl. The precipitate that formed was collected byfiltration and washed with water (2×10 mL) to provide the product as awhite solid (357 mg, 92% yield). ¹H NMR (DMSO-d₆) δ: 12.76 (2H, br),8.05 -8.01 (2H, m), 7.51 -7.41 (4H, m), 6.86 (1H, d, J=3.7 Hz), 6.66(1H, s), 3.23 (2H, t, J=7.2 Hz), 2.81 (2H, t, J=7.4 Hz), 1.80-1.62 (4H,m), 1.51-1.41 (2H, m). Anal. (C₂₀H₂₀N₂O₃S₂.1.5 HCl) C, H, N, S.

Example 2

(a) Methyl 5-(3-hydroxypropynyl)thiophene-2-carboxylate. The titlecompound was prepared by the coupling of methyl5-bromothiophene-2-carboxylate and propargyl alcohol in a manneranalogous to step (a) of Example 1 above. ¹H NMR (CDCl₃) δ: 7.64 (1H, d,J=3.9 Hz), 7.15 (1H, d, J=3.9 Hz), 4.52 (2H, s), 3.88 (3H, s). Anal.(C₉H₈O₃S) C, H, S.

(b) Methyl 5-(3-hydroxypropyl)thiophene-2-carboxylate. The titlecompound was prepared by the reduction of methyl5-(3-hydroxypropynyl)thiophene-2-carboxylate in a manner similar to step(b) of Example 1. ¹H NMR (CDCl₃) δ: 7.64 (1H, d, J=3.8 Hz), 6.82 (1H, d,J=3.8 Hz), 3.86 (3H, s), 3.71 (2H, t, J=6.2 Hz), 2.96 (2H, t, J=7.6 Hz),1.96 (2H, tt, J=6.2, 7.6 Hz). Anal. (C₉H₁₂O₃S) C, H, S.

(c) Methyl 5-(3-bromopropyl)thiophene-2-carboxylate. The title compoundwas prepared from methyl 5-(3-hydroxypropyl)thiophene-2-carboxylate in amanner analogous to step (c) of Example 1. ¹H NMR (CDCl₃) δ: 7.64 (1H,d, J=3.7 Hz), 6.85 (1H, d, J=3.7 Hz), 3.86 (3H, s), 3.43 (2H, t, J=6.4Hz), 3.03 (2H, t, J=7.2 Hz), 2.22 (2H, tt, J=6.4, 7.2 Hz). Anal.(C₉H₁₁O₂SBr) C, H, S, Br.

(d) Methyl5-(3-[(6-phenyl-4(3H)-oxopyrimidin-2-yl)thio]propyl)thiophene-2-carboxylate.The title compound was prepared from 6-phenyl-2-thiouracil and methyl5-(3-bromopropyl)thiophene-2-carboxylate similar to the method describedin step (d) of Example 1. ¹H NMR (DMSO-d₆) δ: 2.10 (2H, m), 3.04 (2H, t,J=7.0 Hz), 3.27 (2H, t, J=7.0 Hz), 3.77 (3H, s), 6.68 (1H, br s), 7.01(1H, d, J=3.7 Hz), 7.47 (3H, m), 7.64 (1H, d, J=3.7 Hz), 7.98 (2H, d,J=7 Hz).

(e)5-(3-[(6-Phenyl-4(3H)-oxopyrimidin-2-yl)thio]propyl)thiophene-2-carboxylicacid (2). The title compound was prepared by hydrolysis of methyl5-(3-[(6-phenyl-4(3H)-oxopyrimidin-2-yl)thio]propyl)thiophene-2-carboxylatein analogy to step (e) of Example 1. ¹H NMR (DMSO-d₆) δ: 2.09 (2H, m),2.99 (2H, t, J=7.0 Hz), 3.27 (2H, t, J=7.0 Hz), 6.68 (1H, s), 6.95 (1H,d, J=3.7 Hz), 7.47 (4H, m), 7.98 (2H, br s). Anal.(C₁₈H₁₆N₂O₃S₂.1.0H₂O.0.1 NaCl) C, H, N, S.

Example 3

(a) 5-(2-Thienyl)thiophene-2-carboxylic acid. A solution of AgNO₃ (8.78g, 51.7 mmol) in 20 mL of water was added to a vigorously stirredsolution of NaOH (4.13 g, 103 mmol) in 20 mL of water. To the resultingslurry was added 2,2′-bisthiophene-5-carboxaldehyde (5.02 g, 25.8 mmol).The reaction mixture was stirred without any external temperatureregulation for 2.5 hours, and then filtered. The filtrate wassubsequently acidified by addition of concentrated HCl. The precipitatethat formed was collected by filtration and washed with water (2×10 mL)to provide the product as a yellow solid (3.64 g, 67% yield). ¹H NMR(DMSO-d₆) δ: 13.17 (1H, br s), 7.65 (1H, d, J=3.9 Hz), 7.62 (1H, dd,J=1.1, 5.2 Hz), 7.48 (1H, dd, J=1.1, 3.7 Hz), 7.34 (1H, d, J=3.9 Hz),7.13 (1H, dd, J=3.7, 5.2 Hz). Anal. (C₉H₆O₂S₂.0.1 H₂) C, H, S.

(b) 2-Acetyl-5-(2-thienyl)thiophene. A 1.5M solution of methyllithium inether (19 mL, 28.5 mmol) was added to a solution of5-(2-thienyl)thiophene-2-carboxylic acid (2.41 g, 11.5 mmol) in 150 mLof ether at −5° C. under an argon atmosphere. The cooling bath wasremoved and the reaction was stirred at ambient temperature fir 90minutes, then poured into 1N HCl (150 mL). The layers were separated andthe aqueous phase was extracted with ether (100 mL). The combinedorganic extracts were washed with saturated NaHCO₃ (150 mL), dried overMgSO₄, and concentrated in vacuo, to give a green solid, which waspurified by flash chromatography. Elution with hexane:EtOAc (85:15)provided the product as a yellow solid (1.89 g, 79% yield). ¹H NMR(DMSO-d₆) δ: 7.89 (1H, d, J=4.0 Hz), 7.64 (1H, dd, J=1.1, 5.1 Hz), 7.52(1H, dd, J=1.1, 3.7 Hz), 7.42 (1H, d, J=4.0 Hz), 7.14 (1H, dd, J=3.7,5.1 Hz), 2.52 (3H, s). Anal. (C₁₀H₈OS₂) C, H, S.

(c) Methyl 3-[5-(2-thienyl)thien-2-yl]-3-oxopropionate. A solution of2-acetyl-5-(2-thienyl)thiophene (1.84 g, 8.8 mmol) in 25 mL of DMF wasadded under an argon atmosphere to a stirred suspension of 60% NaH inoil (777 mg, 19.4 mmol) in 15 mL of DMF at about 0° C. over a 10-minuteinterval. The resulting solution was stirred at 0° C. for about 30minutes preceding addition of dimethylcarbonate (3 mL, 3.21 g, 35.6mmol). The cooling bath was removed and the reaction was stirred atambient temperature for 3 hours, and then poured into water (250 mL) andextracted with ether (100 mL). The aqueous layer was subsequentlyacidified by addition of conc. HCl and extracted with ether (2×100 mL).The combined organic extracts were dried over MgSO₄ and concentrated invacuo, to give a brown gum, which was purified by flash chromatography.Elution with hexane:EtOAc (3:1) provided the product as a yellow solid(2.10 g, ⁸⁹% yield). ¹H NMR (DMSO-d₆) δ: 7.96 (1H, d, J=4.0 Hz), 7.69(1H, dd, J=1.1, 5.1 Hz), 7.56 (1H, dd, J=1.1, 3.7 Hz), 7.46 (1H, d,J=4.0 Hz), 7.16 (1H, dd, J=3.7, 5.1 Hz), 4.13 (2H, s), 3.65 (3H, s).Anal. (C₁₂H₁₀O₃S₂) C, H, S.

(d) 6-[5-(2-Thienyl)thien-2-yl]-4(3H)-oxopyrimidine-2-thiol. Thiourea(883 mg, 11.6 mmol) and a solution of methyl3-[5-(2-thienyl)thien-2-yl]-3-oxopropionate (2.06 g, 7.7 mmol) inethanol (100 mL) were added sequentially to a 1M solution of sodiumethoxide in ethanol (15 mL). The resulting mixture was heated at refluxunder argon for about 24 hours. After removal of the ethanol byconcentration in vacuo, the residue was dissolved in water (120 mL). Theaqueous layer was acidified by addition of conc. HCl and the precipitatethat formed was collected by filtration, and then washed with water(3×30 mL) and with ether (4×30 mL) to provide the product as a yellowsolid (1.04 g, 46% yield). ¹NMR (DMSO-d₆) δ: 12.50 (1H, br s), 12.48(1H, br s), 8.00 (1H, d, J=4.0 Hz), 7.64 (1H, dd, J=1.1, 5.1 Hz), 7.46(1H, dd, J=1.1, 3.7 Hz), 7.41 (1H, d, J=4.0 Hz), 7.14 (1H, dd, J=3.7,5.1 Hz), 6.01 (1H, s).

(e) Methyl5-[5-([6(5-[2-thienyl]thien-2-yl)-4(3H)-oxopyrimidin-2-yl]thio)pentyl]thiophene-2-carboxylate.The title compound was prepared from6-[5-(2-thienyl)thien-2-yl]-4(3H)-oxopyrimidine-2-thiol and methyl5-(5-bromopentyl)thiophene-2-carboxylate in a manner similar to Example1(d). ¹H NMR (DMSO-d₆) δ: 12.68 (1H, br s), 7.82 (1H, d, J=4.0 Hz), 7.57(1H, d, J=3.8 Hz), 7.56 (1H, dd, J=1.1, 5.1 Hz),7.3 (1H, d, J=4.0 Hz),7.36 (1H, dd, J=1.1, 3.7 Hz), 7.10 (1H, dd, J=3.7, 5.1 Hz), 6.92 (1H, d,J=38 Hz), 6.60 (1H, s), 3.75 (3H, s), 3.17 (2H, t, J=7.3 Hz), 2.86 (2H,t, J=7.4 Hz), 1.82-1.66 (4H, m), 1.54-1.45 (2H, m). Anal. (C₂₃H₂₂N₂O₃S₄)C, H, N, S.

(f)5-[5-([6-(5-[2-Thienyl]thien-2-yl)-4(3H)-oxopyrimidin-2-yl]thio)pentyl]thiophene-2-carboxylicacid (3). The title compound was prepared by hydrolysis of methyl5-[5-([6-(5-[2-thienyl]thien-2-yl)4(3H)-oxopyrimidin-2-yl]thio)pentyl]thiophene-2-carboxylatein analogy to Example 1(e). ¹H NMR (DMSO-d₆) δ: 12.82 (1H, broad), 7.81(1H, d, J=4.0 Hz), 7.55 (1H, dd, J=1.1, 5.1 Hz), 7.43 (1H, d, J=3.6 Hz),7.37 (1H, d, J=4.0 Hz), 7.36 (1H, dd, J=1.1, 3.7 Hz), 7.11 (1H, dd,J=3.7, 5.1 Hz), 6.84 (1H, d, J=3.6 Hz), 6.60 (1H, s), 3.16 (2H, t, J=7.3Hz), 2.83 (2H, t, J=7.4 Hz), 1.82-1.70 (4H, m), 1.45-1.45 (2H, m). Anal.(C₂₂H₂₀N₂O₃S₄.1.5 HCl) C, H, N, S.

Example 4

(a) Methyl-4-phenylbenzoylacetate. To a stirred suspension of 60% NaH inoil (4.5 g, 112 mmol) in 30 mL of DMF at RT (room temperature) was addeddropwise 4-acetylbiphenyl (10 g, 51 mmol) dissolved in 50 mL of DMF.After 30 minutes, the mixture was cooled to 0° C. and dimethylcarbonate(27 mL, 331 mmol) was added neat. The mixture was allowed to warm to RTover a 2-hour period and then poured into 1:1 water/saturatedbicarbonate. The aqueous layer was extracted with Et₂O, and afterremoval of the solvent under vacuum, the crude residue was flashchromatographed with 0-40% EtOAc/hexanes to give 12.1 g (94%) of thedesired product as a crystalline solid: ¹H NMR (CDCl₃) δ: 3.78 and 3.83(3H, s), 4.05 and 5.73 (2H, s), 7.38-7.51 (3H, m), 7.62-7.72 (4H, m),7.83 (1H, d, J=8.5 Hz), 8.03 (2H, d, J=8.6 Hz), 12.53 (0.2H, s).

(b) 6-(4-Biphenylyl)-4(3H)-oxopyrimidine-2-thiol. To a stirred solutionof methyl-4-phenylbenzoylacetate (5 g, 19 mmol) and thiourea (2.24 g,29.5 mmol) in 30 mL of EtOH was added 21% by weight NaOEt in EtOH (15mL, 41 mmol). The mixture was heated to reflux under argon for 18 hours.After cooling to RT, the suspension was filtered and washed with EtOHand dissolved in water. The solution was acidified with 2N HCl until thepH was 5, and the resulting suspension was filtered, washed with water,and dried to give 2.4 g (43%) of the desired product as a white solid:¹H NMR (DMSO-d₆) δ: 6.15 (1H, s), 7.41-7.52 (3H, m), 7.72-7.80 (5H, m),8.00 (1H, d, J=8.1 Hz), 12.51 (1H, d, J=8.0 Hz).

(c) Methyl5-[5-([6-(4-biphenylyl)-4(3H)-oxopyrimidin-2-yl]thio)pentyl]-thiophene-2-carboxylate(4). The title compound was prepared from6-(4′-biphenylyl)-4(3H)-oxopyrimidine-2-thiol and methyl5-(5-bromopentyl)thiophene-2-carboxylate in analogy to Example 1(d). ¹HNMR (DMSO-d₆) δ: 1.50 (2H, m), 1.79 (4H, m), 2.85 (2H, t, J=7.0 Hz),3.26 (2H, br s), 3.73 (3H, s), 6.70 (1H, br s), 6.93 (1H, d, J=3.7 Hz(3H, m), 7.59 (1H, d, J=3.7 Hz), 7.70-7.75 (5H, m), 8.13 (1H, d, J=8.1Hz), 12.75 (1H, br s). Anal. (C₂₇H₂₆N₂O₃S₂) C, H, N, S.

Example 5

5-[5-([6-(4-Biphenylyl)-4(3H)-oxopyrimidin-2-yl]thio)pentyl]thiophene-2-carboxylicacid (5). The title compound was prepared by hydrolysis of methyl5-[5-([6-(4′-biphenylyl)-4(3H)-oxopyrimidin-2-yl]thio)pentyl]thiophene-2-carboxylatein analogy to Example 1(e). ¹H NMR (DMSO-d₆) δ: 1.48 (2H, m), 1.67-1.77(4H, m), 2.83 (2H, t, J=7.0 Hz), 3.25 (2H, br s), 6.72 (1H, s), 6.88(1H, d, J=3.7 Hz), 7.38-7.51 (3H, m), 7.70-7.76 (6H, m), 8.14 (2H, d,J=8.5 Hz), 12.80 (1H, br s). Anal. (C₂₆H₂₄N₂O₃S₂.0.7 H₂O) C, H, N, S.

Example 6

(a) Methyl-4-phenoxybenzoylacetate. The title compound was prepared in amanner as described above in Example 4(a) using 4-phenoxyacetophenone togive the desired product as a yellow solid. ¹H NMR (CDCl₃) δ: 3.69 (3H,s), 3.91 (2H, s), 6.94 (2H, d, J=8.5 Hz), 7.01 (2H, d, J=8.1 Hz),7.11-7.20 (2H, m), 7.35 (2H, t, J=7.7 Hz), 7.86 (2H, d, J=8.8 Hz).

(b) 6-(4-Phenoxyphenyl)-4(3H)-oxopyrimidine-2-thiol. The title compoundwas prepared from methyl-4-phenoxybenzoylacetate and thiourea in themanner described in Example 4(b). ¹H NMR (DMSO-d₆) δ: 3.33 (3H, s), 5.97(1H, s), 6.83-7.19 (5H, m), 7.41 (2H, m), 7.92 (2H, d, J=8.8 Hz), 10.40(1H, br s).

(c) Methyl5-[5-([6-(4-phenoxyphenyl)-4(3H)-oxopyrimidin-2-yl]thio)pentyl]-thiophene-2-carboxylate.The title compound was prepared from6-(4′-phenoxyphenyl)-4(3H)-oxopyrimidine-2-thiol and methyl5-(5-bromopentyl)thiophene-2-carboxylate in a manner as described inExample 1(d). ¹H NMR (DMSO-d₆) δ: 1.47 (2H, m), 1.72 (4H, m), 2.83 (2H,t, J=7.4 Hz), 3.22 (2H, t, J=7.4 Hz), 3.76 (3H, s), 6.60 (1H, br s),6.90 (1H, d, J=3.7 Hz), 7.00 (2H, d, J=8.8 Hz), 7.08 (2H, d, J=7.7 Hz),7.20 (2H, t, J=7.4 Hz), 7.43 (1H, t, J=7.7 Hz), 7.58 (1H, d, J=3.7 Hz),8.06 (1H, d, J=8.8 Hz), 12.72 (1H, br s). Anal. (C₂₇H₂₆N₂O₄S₂) C, H, N,S.

(d)5-[5-([6-(4-Phenoxypheuyl)-4(3H)-oxopyrimidin-2-yl]thio)pentyl]thiophene-2-carboxylicacid (6). The title compound was prepared by hydrolysis of methyl5-[5-([6-(4′-phenoxyphenyl)-4(3H)-oxopyrimidin-2-yl]thio)pentyl]thiophene-2-carboxylatein a manner like that previously described in Example 1(e). ¹H NMR(DMSO-d₆) δ: 1.44 (2H, m), 1.72 (4H, m), 2.76 (2H, t, J=8.1 Hz), 3.21(2H, t, J=7.0 Hz), 6.61 (1H, br s), 6.82 (1H, d, J=3.7 Hz), 7.02 (2H, d,J=8.8 Hz) 7.09 (2H, d, J=7.4 Hz), 7.20 (1H, t, J=7.4 Hz), 7.43 (3H, m),8.06 (2H, d, J=8.8 Hz). Anal. (C₂₆H₂₄N₂O₄S₂.0.8H₂O) C, H, N, S.

Example 7

(a) Methyl-4-morpholinylbenzoylacetate. The title compound was preparedas described above in Example 4(b), except using4-morpholinylacetophenone to give the desired product as a yellow solid.¹H NMR (CDCl₃) δ: 3.7 (4H, m), 3.78 (3H, s), 3.90 (4H, m), 3.97 (2H, s),6.92 (2H, d, J=9.2 Hz), 7.91 (2H, d, J=8.8 Hz).

(b) 6(4-Morpholinylphenyl)-4(3H)-oxopyrimidine-2-thiol. The titlecompound was prepared using the procedure described above in Example4(b), except using methyl-4-morpholinylbenzoylacetate and thiourea togive the desired product as a light brown solid. ¹H NMR (DMSO-d₆) δ:3.25 (4H, m), 3.72 (4H, m), 6.01 (1H, s), 6.98 (2H, d, J=9.2 Hz), 7.63and 7.79 (2H, d, J=8.8 Hz), 12.20 (1H, s), 12.38 (1H, s).

(c) Methyl5-[5-([6-(4-morpholinylphenyl)-4(3H)-oxopyrimidin-2-yl]thio)pentyl]-thiophene-2-carboxylate.The title compound was prepared from6-(4′-morpholinylphenyl)-4(3H)-oxopyrimidine-2-thiol and methyl5-(5-bromopentyl)thiophene-2-carboxylate analogously to Example 1(d). ¹HNMR (DMSO-d₆) δ: 1.50 (2H, m), 1.68 (4H, m), 2.82 (2H, t, J=7.4 Hz),3.19 (4H, br s), 3.71 (4H, br s), 3.78 (3H, s), 6.48 (1H, br s), 6.93(3H, m), 7.61 (1H, d, J=4.0 Hz), 7.92 (1H, d, J=9.6 Hz).

(d)5-[5-([6-(4-Morpholinylphenyl)-4(3H)-oxopyrimidin-2-yl]thio)pentyl]-thiophene-2-carboxylicacid (7). The title compound was prepared by hydrolysis of methyl5-[5-([6-(4′-morpholinylphenyl)-4(3H)-oxopyrimidin-2-yl]thio)pentyl]thiophene-2-carboxylateas previously described in Example 1(e). ¹H NMR (DMSO-d₆) δ: 1.45 (2H,m), 1.75 (4H, m), 2.83 (2H, t, J=7.0 Hz), 3.20 (4H, m), 3.75 (4H, m),6.51 (1H, br s), 6.92 (3H, m), 7.54 (1H, d, J=3.7 Hz), 7.93 (2H, d,J=8.8 Hz), 12.68 (2H, br s). Anal. (C₂₄H₂₇N₃O₄S₂.0.9H₂O) C, H, N, S.

Example 8

(a) 5-Cyano-6-[5-(2-thienyl)thien-2-yl]-4(3H)-oxopyrimidine-2-thiol. Toa stirred solution of 2,2′-bisthiophene-5-carboxaldehyde (1.9 g, 10mmol) in ethanol (50 mL) were added, sequentially, ethyl cyanoacetate(1.13 g, 10 mmol), thiourea (0.76 g, 10 mmol), and K₂CO₃ (1.4 g, 10mmol). The reaction mixture was heated at reflux for 16 hours. Aftercooling to room temperature, the precipitate was collected byfiltration, washed with ethanol (2×20 mL), and then dissolved in warmwater (1 L). The aqueous solution was allowed to cool to roomtemperature, and then acidified to pH 5 by addition of acetic acid. Theprecipitate that formed was collected by filtration and washed withwater (2×20 mL) to provide the product as a yellow solid (1.6 g, 50%yield). ¹H NMR (DMSO-d₆) δ: 12.99 (1H, s), 7.97 (1H, d, J=4.1 Hz), 7.68(1H, dd, J=1.0, 5.1 Hz), 7.54 (1H, dd, J=1.0, 3.6 Hz), 7.50 (1H, d,J=4.1 Hz), 7.17 (1H, dd, J=3.6, 5.1 Hz).

(b) Methyl5-[5([5-cyano-6-(5-[2-thienyl]thien-2-yl)-4(3H)-oxopyrimidin-2-yl]thio)pentyl]thiophene-2-carboxylate.The title compound was prepared from5-cyano-6-[5-(2-thienyl)thien-2-yl]-4(3H)-oxopyrimidine-2-thiol andmethyl 5-(5-bromopentyl)thiophene-2-carboxylate in a manner like thatpreviously described in Example 1(d). ¹H NMR (DMSO-d₆) δ: 8.19 (1H, d,J=4.2 Hz), 7.65 (1H, dd, J=1.0, 5.0 Hz), 7.56 (1H, d, J=3.7 Hz), 7.53(1H, d, J=4.2 Hz), 7.47 (1H, dd, J=1.0, 3.6 Hz), 7.15 (1H, dd, J=3.6,5.0 Hz), 6.91 (1H, d, J=3.7 Hz), 3.75 (3H, s), 3.23 (2H, t, J=7.4 Hz),2.86 (2H, t, J=7.3 Hz), 1.84-1.66 (4H, m), 1.57-1.46 (2H, m). Anal.(C₂₄H₂₁N₃O₃S₄.0.33 H₂O) C, H, N, S.

(c)[5-([5-Cyano-6-(5-[2-thienyl]thien-2-yl)-4(3H)-oxopyrimidin-2-yl]thio)pentyl]thiopbene-2-carboxylicacid (8). The title compound was prepared by hydrolysis of methyl5-[5-([5-cyano-6-(5-[2-thienyl]thien-2-yl)-4(3H)-oxopyrimidin-2-yl]thio)pentyl]thiophene-2-carboxylatein a manner like that described in Example 1(e). ¹H NMR (DMSO-d₆) δ:8.19 (1H, d, J=4.2 Hz), 7.66 (1H, dd, J=1.0, 5.1 Hz), 7.53 (1H, d, J=4.2Hz), 7.50 (1H, d, J=3.7 Hz), 7.48 (1H, dd, J=1.0, 3.6 Hz), 7.15 (1H, dd,J=3.6, 5.1 Hz), 6.88 (1H, d, J=3.7 Hz), 3.23 (2H, t, J=7.2 Hz), 2.85(2H, t, J=7.4 Hz), 1.75-1.56 (4H, m), 1.47-1.36 (2H, m). Anal.(C₂₃H₁₉N₃O₃S₄.0.5 H₂O) C, H, N, S.

Example 9

(a) Diethyl N-(5-bromothien-2-yl)glutamate. To a stirred solution of5-bromothiophene-2-carboxylic acid (15.53 g, 75 mmol),1-hydroxybenzotriazole (10.81 g, 80 mmol), L-glutamic acid diethyl esterhydrochloride (19.18 g, 80 mmol), and diisopropylethylamine (14 mL,10.39 g, 80 mmol) in DMF (75 mL) was added1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (15.34 g, 80mmol). The resulting solution was stirred under argon at ambienttemperature for 18 hours, poured into brine (300 mL), diluted with water(300 mL), and extracted with ether (3×150 mL). The combined organicextracts were washed with water (2×150 mL), dried over MgSO₄, andconcentrated in vacuo, to give a yellow gum which was purified by flashchromatography. Elution with hexane:EtOAc (2:1) provided the product asa yellow syrup (26.81 g, 91% yield). ¹H NMR (CDCl₃) δ: 7.29 (1H, d,J=3.7 Hz), 7.04 (1H, d, J=3.7 Hz), 6.98 (1H, d, J=7.7 Hz), 4.69 (1H,ddd, J=4.8, 7.7, 12.5 Hz), 4.23 (2H, q, J=7.2 Hz), 4.12 (2H, q, J=7.2Hz), 2.54-2.37 (2H, m), 2.33-2.22 (1H, m), 2.18-2.04 (1H, m), 1.30 (3H,t, J=7.2 Hz), 1.24 (3H, t, J=7.2 Hz). Anal. (C₁₄H₁₈NO₅SBr) C, H, N, S,Br.

(b) Diethyl N-[5-(5-hydroxy-1-pentynyl)theno-2-yl]glutamate. The titlecompound was prepared from the coupling of diethylN-(5-bromothien-2-yl)glutamate and 4-pentyn-1-ol as in Example 1(d). ¹HNMR (CDCl₃) δ: 1.2-1.32 (6H, m), 1.84-1.88 (2H, m), 2.1-2.48 (5H, m),2.57 (2H, t, J=7.00 Hz), 3.80 (2H, t, J=6.25 Hz), 4.11 (2H, q, J=7.00Hz), 4.22 (2H, q, J=7.00 Hz), 4.7 (1H, m), 6.9 (1H, d, J=5 Hz), 7.04(1H, d, J=3.68 Hz), 7.37 (1H, d, J=4.1 Hz).

(c) Dietbyl N-[5-(5-hydroxypentyl)theno-2-yl]glutamate. The titlecompound was prepared by the reduction of diethylN-[5-(5-hydroxy-1-pentynyl)theno-2-yl]glutamate as previously describedin Example 1(e). ¹H NMR (CDCl₃) δ: 1.2-1.32 (6H, m), 1.4-1.77 (10H, m),2.07-2.54 (5H, m), 2.85 (2H, t, J=7.4 Hz) 3.66 (2H, t, J=6.25 Hz), 4.13(2H, q, J=7.00 Hz), 4.23 (2H, q, J=7.00 Hz), 4.76 (1H, m), 6.76 (2H, m),7.38 (1H, d, J=3.3 Hz).

(d) Diethyl N-[5-(5bromopentyl)theno-2-yl]glutamate. The title compoundwas prepared from diethyl N-[5-(5-hydroxypentyl)theno-2-yl]glutamatelike as described in Example 1(c). ¹H NMR (CDCl₃) δ: 1.23-1.35 (6H, m),1.5-1.94 (10H, m), 2.07-2.53 (5H, m), 2.87 (2H, t, J=7.4 Hz), 3.43 (2H,t, J=6.25 Hz), 4.14 (2H, q, J=7.00 Hz), 4.25 (2H, q, J=7.00 Hz), 4.76(1H, m), 6.79 (2H, m), 7.40 (1H, d, J=3.68 Hz).

(e) DiethylN-(5-[5-([6-phenyl-4(3H)-oxopyrimidin-2-yl]thio)pentyl]theno-2-yl)glutamate.The title compound was prepared from 6-phenyl-2-thiouracil and diethylN-[5-(5-bromopentyl)theno-2-yl]glutarnate in like manner to Example1(d). ¹H NMR (DMSO-d₆) δ: 1.16 (6H, q, J=7.0 Hz), 1.4-2.2 (10H, m), 2.39(2H, t, J=8.1 Hz), 2.81 (2H, t, J=8.1 Hz), 3.33 (2H, t, J=6.25 Hz),4.04-4.18 (4H, m), 4.40 (1H, m), 6.68 (1H, m), 6.86 (1H, t, J=3.4 Hz),7.46 (3H, br s), 7.67 (1H, d, J=3.4 Hz), 8.03 (2H, br s), 8.61 (1H, d,J=12.78 (1H, br s).

(f)N-(5-[5-([6-Phenyl-4(3H)-oxopyrimidin-2-yl]thio)pentyl]theno-2-yl)glutamicacid (9). The title compound was prepared by hydrolysis of diethylN-(5-[5-([6-phenyl-4(3H)-oxopyrimidin-2-yl]thio)pentyl]theno-2-yl)glutamatein analogy to Example 1(e). ¹H NMR (DMSO-d₆) δ: 1.4-2.2 (10H, m), 2.33(2H, t, J=7.3 Hz), 2.81 (2H, t, J=7.0 Hz), 3.23 (2H, t, J=7.0 Hz), 4.31(1H, m), 6.67 (1H, s), 6.86 (1H, t, J=3.7 Hz), 7.45 (3H, br s), 7.67(1H, d, J=3.7 Hz), 8.03 (2H, m), 8.48 (1H, d, J=7.7 Hz) 12.60 (3H, brs). Anal. (C₂₅H₂₇N₃O₆S₂.0.5H₂O) C, H, N, S.

Example 10

(a) DiethylN-(5-[5-([5-cyano-6-(5-[2-thienyl]thien-2-yl)-4(3H)-oxopyrimidin-2-yl]thio)pentyl]theno-2-yl)glutamate.The title compound was prepared from5-cyano-6-[5-(2-thienyl)thien-2-yl]-4(3H)-oxopyrimidine-2-thiol anddiethyl N-[5-(5-bromopentyl)theno-2-yl]glutamate in analogy to Example1(d). ¹H NMR (CDCl₃) δ: 8.41 (1H, d, J=4.2 Hz), 7.36 (1H, dd, J=1.1, 5.0Hz), 7.35 (1H, d, J=3.7 Hz), 7.34 (1H, dd, J=1.1, 3.6 Hz), 7.29 (1H, d,J=4.2 Hz), 7.08 (1H, dd, J=3.6, 5.0 Hz), 6.82 (1H, d , J=6.73 (1H, d,J=3.7 Hz), 4.74 (1H, ddd, J=4.8, 7.8, 11.6 Hz), 4.24 (2H, q, J=7.1 Hz),4.11 (2H, q, J=7.1 Hz), 3.32 (2H, t, J=7.0 Hz), 2.87 (2H, t, J=7.2 Hz),2.56-2.24 (3H, m), 2.18-2.08 (1H, m), 1.93-1.72 (4H, m), 1.63-1.52 (2H,m), 1.30 (3H, t, J=7.1 Hz), 1.23 (3H, t, J=7.1 Hz).

(b)N-(5-[5-([5-Cyano-6-(5-[2-thienyl]thien-2-yl)-4(3H)-oxopyrimidin-2-yl]thio)pentyl]theno-2-yl)glutamicacid (10). The title compound was prepared by hydrolysis of diethylN-(5-[5-([5-cyano-6-(5-[2-thienyl]thien-2-yl)-4(3H)-oxopyrimidin-2-yl]thio)pentyl]theno-2-yl)glutamatein a manner akin to Example 1(e). ¹H NMR (DMSO-d₆) δ: 8.45 (1H, d ,J=7.7 Hz), 8.18 (1H, d, J=4.2 Hz), 7.65 (1H, dd, J=1.1, 5.0 Hz), 7.64(1H, d, J=3.7 Hz), 7.53 (1H, d, J=4.2 Hz), 7.48 (1H, dd, J=1.1, 3.6 Hz),7.15 (1H, dd, J=3.6, 5.0 Hz), 6.85 (1H, d, J=3.7 Hz), 4.33 (1H, ddd,J=4.9, 7.7, 9.3 Hz), 3.21 (2H, t, J=7.4 Hz), 2.83 (2H, t, J=7.2 Hz),2.32 (2H, t, J=7.5 Hz), 2.12-1.89 (2H, m), 1.85-1.66 (4m), 1.57-1.46(2H, m). Anal. (C₂₈H₂₆N₄O₆S₄.1.0 H₂O) C, H, N, S.

Example 11

(a) Diethyl N-[5-(4-hydroxy-1-butynyl)theno-2-yl]glutamate. The titlecompound was prepared from the coupling of diethylN-(5-bromothien-2-yl)glutamate and 3-butynyl-1-ol as previouslydescribed for Example 1(a). ¹H NMR (CDCl₃) δ: 7.38 (1H, d, J=40 Hz),7.08 (1H, d, J=4.0 Hz), 6.96 (1H, d, J=7.4 Hz), 4.71 (1H, ddd, J=4.8,7.4, 12.4 Hz), 4.23 (2H, q, J=7.0 Hz), 4.12 (2H, q, J=7.0 Hz), 3.83 (2H,t, J=6.3 Hz), 2.72 (2H, t, J=6.3 Hz), 2.55-2.22 (3H, m), 2.18-2.06 (1H,m), 1.30 (3H, t, J=7.0 Hz), 1.23 (3H, t, J=7.0 Hz). Anal. (C₁₈H₂₃NO₆S)C, H, N, S.

(b) Diethyl N-[5-(4-hydroxybutyl)theno-2-yl]glutamate. The titlecompound was prepared by the reduction of diethylN-[5-(4-hydroxy-1-butynyl)theno-2-yl]glutamate as previously describedfor Example 1(b). ¹H NMR (CDCl₃) δ: 7.39 (1H, d, J=3.7 Hz), 6.77 (1H, d,J=7.7 Hz), 6.76 (1H, d, J=3.7 Hz), 4.73 (1H, ddd, J=5.1, 7.7, 12.1 Hz),4.23 (2H, q, J=7.0 Hz), 4.11 (2H, q, J=7.0 Hz), 3.68 (2H, t, J=6.4 Hz),2.87 (2H, t, J=7.3 Hz), 2.53-2.23 (3H, m), 2.17-2.04 (1H, m), 1.83-1.71(2H, m), 1.68-1.61 (2H, m), 1.30 (3H, t, J=7.0 Hz), 1.23 (3H, t, J=7.0Hz). Anal. (C₁₈H₂₇NO₆S) C, H, N, S.

(c) Diethyl N-[5-(4-bromobutyl)theno-2-yl]glutamate. The title compoundwas prepared from diethyl N-[5-(4-hydroxybutyl)theno-2-yl]glutamate aspreviously described for Example 1(c). ¹H NMR (CDCl₃) δ: 7.39 (1H, d,J=3.7 Hz), 6.79 (1H, d, J=7.7 Hz), 6.77 (1H, d, J=3.7 Hz), 4.73 (1H,ddd, J=4.8, 7.7, 8.1 Hz), 4.23 (2H, q, J=7.0 Hz), 4.11 (2H, q, J=7.0Hz), 3.42 (2H, t, J=6.3 Hz), 2.86 (2H, t, J=7.5 Hz), 2.53-2.23 (3H, m),2.17-2.04 (1H, m), 1.95-1.81 (4H, m), 1.30 (3H, t, J=7.0 Hz), 1.23 (3H,t, J=7.0 Hz). Anal. (C₁₈H₂₆NO₅SBr) C, H, N, S, Br.

(d) DiethylN-(5-[4-([6-phenyl-4(3H)-oxopyrimidin-2-yl]thio)butyl]theno-2-yl)glutamate.The title compound was prepared from 6-phenyl-2-thiouracil and diethylN-[5-(4-bromobutyl)theno-2-yl]glutamate as described above for Example1(d). ¹H NMR (DMSO-d₆) δ: 1.16 (6H, m), 1.8-2.2 (8H, m), 2.38 (2H, t,J=8.5 Hz), 2.89 (2H, d, J=8.5 Hz) 3.34 (2H, br s), 4.00-4.13 (4H, m),4.43 (1H, m), 6.70 (1H, m), 6.87 (1H, t, J=3.7 Hz), 7.48 (3H, br s),7.66 (1H, d, J=3.7 Hz), 8.03 (2H, br s), 8.60 (1H, d, J=7.6 Hz), 12.80(1H, br s).

(e)N-(5-[5-([6-Phenyl-4(3H)-oxopyrimidin-2-yl]thio)butyl]thio2-yl)glutamicacid (11). The title compound was prepared by hydrolysis of diethylN-(5-[4-([⁶-phenyl-4(3H)-oxopyrimidin-2-yl]thio)butyl]theno-2-yl)glutamateas described above for Example 1(e). ¹H NMR (DMSO-d₆) δ: 1.8-2.2 (8H,m), 2.33 (2H, t, J=6.24 Hz), 2.86 (2H, br s) 3.33 (2H, br s), 4.00-4.13(4H, m), 4.35 (1H, m), 6.68 (1H, m), 6.87 (1H, t, J=3.7 Hz), 7.49 (3H,br s), 7.66 (1H, d, J=3.7 Hz), 8.03 (2H, br s), 8.49 (1H, d, J=8.1 Hz),12.60 (3H, br s). Anal. (C₂₄H₂₅N₃O₆S₂.0.1H₂O) C, H, N, S.

Example 12

(a) 2-[2-(t-Butyldimethylsilyloxy)ethyl]thiophene. To a stirred solutionof 2-(2-thienyl)ethanol (75.26 g, 0.6 mol) in methylene chloride (800mL) were added t-butyldimethylchlorosilane (97.34 g, 0.6 mol),triethylamine (96 mL, 69.69 g, 0.7 mol) and 4-(dimethylamino)pyridine(1.22 g, 0.01 mol) at 0° C. The cooling bath was removed and thereaction was stirred at ambient temperature overnight. The precipitate(triethylamine hydrochloride) was removed by filtration, and thefiltrate was washed sequentially with water (400 mL), 0.5N HCl (400 mL),and brine (400 mL). The aqueous layers were combined and extracted withCH₂Cl₂(400 mL). The combined organic extracts were dried over MgSO₄ andconcentrated in vacuo, to provide the product as a yellow oil (139.55 g,98% yield). ¹H NMR (CDCl₃) δ: 7.13 (1H, d, J=5.1 Hz), 6.92 (1H, dd,J=3.3, 5.1 Hz), 6.83 (1H, d, J=3.3 Hz), 3.82 (2H, t, J=6.7 Hz), 3.03(2H, t, J=6.7 Hz), 0.89 (9H, s), 0.03 (6H, s). Anal. (C₁₂H₂₂OSSi) C, H,S.

(b) 5-[2-(t-Butyldimethylsilyloxy)ethyl]thiophene-2-carboxylic acid,dietbyl N-(5-[2-(t-butyldimetlylsilyloxy)ethyl]theno-2-yl)glutamate anddiethyl N-[5-(2-hydroxyethyl)theno-2-yl]glutamate. A 2.5M solution ofn-butyllitium in hexane (300 mL, 0.75 mol) was added to a solution of2-[2-(t-butyldimethylsilyloxy)ethyl]thiophene (139.55 g, 0.58 mol) inTHF (1L) under an argon atmosphere at −75° C. The resulting reactionmixture was stirred at −75° C. for about an hour, then warmed to −20° C.Dry CO₂ was then bubbled through the reaction for 90 minutes. The crudemixture was poured into a mixture of ice (2.5 kg) and saturated NH₄Cl(700 mL). The layers were separated, and the aqueous phase was extractedwith ether (700 mL) and EtOAc (700 mL). The combined organic extractswere dried over Na₂SO₄ and concentrated in vacuo, to give an orangesolid (180 g), which was used without further purification.

The crude 5-[2-(t-butyldimethylsilyloxy)ethyl]thiophene-2-carboxylicacid was subsequently coupled with L-glutamic acid diethyl esterhydrochloride as described in Example 9(a), to give a brown syrup (200g), which was used without further purification.

The crude diethylN-(5-[2-(t-butyldimetlylsilyloxy)ethyl]theno-2-yl)glutamate wasdissolved in THF (800 mL) and combined with a 1.0M solution oftetrabutylammonium fluoride (500 mL, 0.5 mol). The resulting solutionwas stirred at ambient temperature overnight, then poured into water (2L) and extracted with EtOAc (3×500 mL). The combined organic extractswere washed with brine (600 mL), dried over Na₂SO₄ and concentrated invacuo, to give a brown gum, which was purified by flash chromatography.Elution with hexane:EtOAc (2:1) provided the product as an orange syrup(99.42 g, 48% yield). ¹H NMR (CDCl₃) δ: 7.41 (1H, d, J=3.7 Hz), 6.88(1H, d, J=7.7 Hz), 6.85 (1H, d, J=3.7 Hz), 4.73 (1H, ddd, J=4.8, 7.7,12.5 Hz), 4.23 (2H, q, J=7.0 Hz), 4.11 (2H, q, J=7.0 Hz), 3.88 (2H, t,J=6.3 Hz), 3.07 (2H, t, J=6.3 Hz), 2.55-2.23 (3H, m), 2.16-2.02 (1H, m),1.30 (3H, t, J=7.0 Hz), 1.23 (3H, t, J=7.0 Hz). Anal. (C₁₆H₂₃NO₆S) C, H,N, S.

(c) Diethyl N-[5-(2-bromoethyl)theno-2-yl]glutamate. The title compoundwas prepared from diethyl N-[5-(2-hydroxyethyl)theno-2-yl]glutamate aspreviously described in Example 1(c). ¹H NMR (CDCl₃) δ: 7.40 (1H, d,J=3.7 Hz), 6.90 (1H, d, J=8.1 Hz), 6.86 (1H, d, J=3.7 Hz), 4.73 (1H,ddd, J=5.1, 8.1, 12.8 Hz), 4.23 (2H, q, J=7.0 Hz), 4.10 (2H, q, J=7.0Hz), 3.56 (2H, t, J=7.2 Hz), 3.35 (2H, t, J=7.2 Hz), 2.53-2.22 (3H, m),2.16-2.06 (1H, m), 1.29 (3H, t, J=7.0 Hz), 1.22 (3H, t, J=7.0 Hz). Anal.(C₁₆H₂₂NO₅SBr) C, H, N, S, Br.

(d) DiethylN-(5-[2-([6-phenyl4(3H)-oxopyrimidin-2-yl]thio)ethyl]theno-2-yl)glutamate.The title compound was prepared from 6-phenyl-2-thiouracil and diethylN-[5-(2-bromoethyl)theno-2-yl]glutamate generally as described inExample 1(d). ¹H NMR (DMSO-d₆) δ: 1.15 (t, 3H, J=7.0 Hz), 1.18 (t, 3H,J=7.0 Hz), 1.97 (m, 2H), 2.41 (t, 2H, J=7.3 Hz), 3.28 (t, partiallyobscured by H₂O, 2H, J=7.3 Hz), 3.50 (t, 2H, J=7.3 Hz), 4.03 (q, 2H,J=7.0 Hz), 4.10 (q, 2H, J=7.0 Hz), 4.35 (m, 1H), 6.70 (s, 1H), 7.00 (d,1H, J=3.7 Hz), 7.47 (m, 3H), 7.72 (d, 1H, J=3.7 Hz), 8.08 (d, 2H, J=8.1Hz), 8.67 (d, 1H, J=7.3 Hz), 12.80 (br s, 1H). Anal. (C₂₆H₂₉N₃O₆S₂) C,H, N, S.

(e)N-(5-[2-([6-Phenyl-4(3H)-oxopyrimidin-2-yl]thio)ethyl]theno-2-yl)glutamicacid (12). The title compound was prepared by hydrolysis of diethylN-(5-[2-([6-phenyl-4(3H)-oxopyrimidin-2-yl]thio)ethyl]theno-2-yl)glutamategenerally as described in Example 1(e). ¹H NMR (DMSO-d₆) δ: 1.86-2.10(m, 2H), 2.33 (m, 2H), 3.28 (t, partially obscured by H₂O, 2H, J=7.3Hz), 3.51 (t, 2H, J=7.3 Hz), 4.34 (m, 1H), 6.71 (s, 1H), 6.99 (d, 1H,J=3.3 Hz), 7.47 (m, 3H), 7.72 (d, 1H, J=3.7 Hz), 8.09 (d, 2H, J=7.0 Hz),8.56 (d, 1H, J=7.7 Hz), 12.6 (br s, 2H). Anal. (C₂₂H₂₁N₃O₆S₂.0.8H₂O) C,H, N, S.

Example 13

(a) DiethylN-(5-[3-([6-phenyl-4(3H)-oxopyrimidin-2-yl]thio)propyl]theno-2-yl)glutamate.The title compound was prepared from 6-phenyl-2-thiouracil and diethylN-[5-(3-bromopropyl)theno-2-yl]glutamate as described above in Example1(d). ¹H NMR (DMSO-d₆) δ: 1.14 (t, 3H, J=7.0 Hz), 1.16 (t, 3H, J=7.0Hz), 1.95-2.10 (m, 4H), 2.40 (t, 2H, J=7.3 Hz), 2.97 (t, 2H, J=7.3 Hz),3.26 (t, 2H, J=7.3 Hz), 4.01 (q, 2H, J=7.0 Hz), 4.08 (q, 2H, J=7.3 Hz),4.01 (q, 2H, J=7.0 Hz), 4.08 (q, 2H, J=7.0 Hz), 4.36 (m, 1H), 6.67 (s,1H), 6.94 (d, 1H, J=3.7 Hz), 7.47 (m, 3H), 7.70 (d, 1H, J=3.7 Hz), 7.98(m, 2H), 8.64 (d, 1H, J=7.7 Hz), 12.75 (br s, 1H). Anal.(C₂₇H₃₁N₃O₆S₂.0.2H₂O) C, H, N, S.

(b)N-(5-[3-([6Phenyl-4(3H)-oxopyrimidin-2-yl]thio)propyl]theno-2-yl)glutamicacid (13). The title compound was prepared by hydrolysis of diethylN-(5-[3-([6-phenyl-4(3H)-oxopyrimidin-2-yl]thio)propyl]theno-2-yl)glutamateas described above in Example 1(e). ¹H NMR (DMSO-d₆) δ: 1.90 (m, 1H),2.06 (m, 3H), 2.32 (t, 2H, J=7.3 Hz), 2.97 (t, 2H, J=7.3 Hz), 3.27 (t,partially obscured by H₂O, 2H, J=7.3 Hz), 4.32 (m, 1H), 6.67 (s, 1H),6.93 (d, 1H, J=3.7 Hz), 7.47 (m, 3H), 7.70 (d, 1H, J=3.7 Hz), 7.98 (m,2H), 8.51 (d, 1H, J=7.7 Hz), 12.60 (br s, 2H). Anal.(C₂₃H₂₃N₃O₆S₂.0.5H₂O) C, H, N, S.

Example 14

N-(4-Fluorophenyl)4-[([5-cyano-6-(5-[2-thienyl]thien-2-yl)-4(3H)-oxopyrimidin-2-yl]thio)methyl]benzenesulfonamide (14). The title compound was prepared from5-cyano-6-[5-(2-thienyl)thien-2-yl]-4(3H)-oxopyrimidine-2-thiol andN-(4-fluorophenyl) 4-(bromomethyl) benzenesulfonamide generally asdescribed in Example 1(d). ¹H NMR (DMSO-d₆) δ: 4.44 (s, 2H), 7.01-7.13(m, 5H), 7.43-7.64 (m, 8H), 8.07 (d, 1H, J=3.7 Hz), 10.21 (s, 1H), 13.8(br s, 1H). Anal. (C₂₆H₁₇N₄O₃S₄F.1.4H₂O) C, H, N.

Example 15

(a) (4-Fluorophenyl)methylamine. A stirred solution of 4-fluoroaniline(5.01 g, 45.1 mmol), methyliodide (3.09 mL, 49.60 mmol) andN,N-diisopropylethylamine (9.4 mL, 54.10 mmol) in 50 mL of DMF washeated at 70° C. for 1 hour. The cooled reaction mixture was poured intoH₂O and extracted with EtOAc (2 times). The combined organic layer waswashed with saturated NaCl solution (3×), dried over MgSO₄, andconcentrated at reduced pressure. After chromatography on silica (7:1hexanes/EtOAc), the title compound was isolated as an amber liquid(46%). ¹H NMR (CDCl₃) δ: 2.85 (s, 3H), 6.77 (m, 2H), 6.95 (m, 2H). Anal.(C₇H₈NF) C, H, N.

(b) N-(4-Fluorophenyl)-N-methyl 4-(bromomethyl)benzenesulfonamide. To astirred solution of (4-fluorophenyl)methylamine (0.42 g, 3.36 mmol) andN,N-diisopropylethylamine (0.64 mL, 3.69 mmol) in 20 mL of CH₂Cl₂ wasadded 4-(bromomethyl)benzenesulfonyl chloride (0.905 g, 3.36 mmol).After 1 hour at RT the reaction mixture was poured into 0.5N HCl andextracted with CH₂Cl₂ (2×). The combined organic layers were washed withsaturated NaCl solution, dried over MgSO₄ and the solvent removed underreduced pressure. After chromatography on silica (5:1 hexanes/EtOAc),the title compound was isolated as an oil, which solidified on standing(76%). ¹H NMR(CDCl₃) δ: 3.16 (s, 3H), 4.49 (s, 2H), 7.04 (m, 4H), 7.50(m, 4H).

(c) N-(4-Fluorophenyl)-N-methyl4-[([5cyano-6-(5-[2-thienyl]thien-2-yl)-4(3H)-oxopyrimidin-2-yl]thio)methyl]benzenesulfonamide(15). The title compound was prepared from5-cyano-6-[5-(2-thienyl)thien-2-yl]-4(3H)-oxopyrimidine-2-thiol andN-(4-fluorophenyl)-N-methyl 4-(bromomethyl)benzenesulfonamide as inExample 1(d). ¹H NMR (DMSO-d₆) δ: 3.04 (s, 3H), 4.61 (s, 2H), 7.05 (m,4H), 7.14 (m, 2H), 7.49 (m, 4H), 7.65 (d, 1H, J=5.2 Hz), 7.69 (d, 2H,J=8.1 Hz), 8.17 (d, 1H, J=3.7 Hz). Anal. (C₂₇H₁₉N₄O₃S₄F.1.6H₂O) C, H, N,S.

Example 16

N-(2-Methylquinolin-6-yl)4-[([5-cyan-6-(5-[2-thienyl]thien-2yl)-4(3H)-oxopyrimidin-2-yl]thio)methyl]benzenesulfonamide(16). The title compound was prepared from5-cyano-6-[5-(2-thienyl)thien-2-yl]-4(3H)-oxopyrimidine-2-thiol andN-(2-methylquinolin-6-yl) 4-(bromomethyl)benzenesulfonamide as inExample 1(d). ¹H NMR (DMSO-d₆) δ: 2.56 (s, 3H), 4.28 (s, 2H), 7.10 (d,1H, J=3.7 Hz with fine splitting), 7.30 (d, 1H, J=8.5 Hz), 7.35 (d, 1H,J=4.0 Hz), 7.40-7.58 (m, 6H), 7.71 (d, 2H, J=8.5 Hz), 7.76 (d, 1H, J=9.2Hz), 7.98 (d, 1H, J=4.0 Hz), 8.08 (d, 1H, J=8.5 Hz), 10.58 (s, 1H).Anal. (C₃₀H₂₁N₅O₃S₄.0.3CH₂Cl₂.1.4H₂O) C, H, N, S.

Example 17

(a) Methyl 4-amino-3-methylbenzoate. To a solution of3-methyl-4-nitrobenzoic acid (6.50 g, 33.3 mmol) in 100 mL of MeOH wasadded 700 mg of 5% Pd/C. The mixture was hydrogenated at 48 psi H₂ for24 hours, the catalyst was removed by suction filtration, and thefiltrate was concentrated under reduced pressure. The title compound wasobtained as a white solid (99%). ¹H NMR (CDCl₃) δ: 2.19 (s, 3H), 3.85(s, 3H), 4.20 (br s, 2H), 6.65 (d, 2H, J=8.1 Hz), 7.72 (d, 2H, J=8.1Hz), 7.76 (s, 1H). Anal. (C₉H₁₁NO₂) C, H, N.

(b) Methyl 4-(chlorosulfonyl)-3-methylbenzoate. The procedure describedin Chem. Ber. 90 (1957), 841, was used to convert methyl4-amino-3-methylbenzoate to the title sulfonyl chloride in 56% yield. ¹HNMR (CDCl₃) δ: 2.84 (s, 3H), 3.98 (s, 3H), 8.07 (d, 1H, J=8.1 Hz), 8.08(s, 1H), 8.14 (d, 1H, J=8.1 Hz). Anal. (C₉H₉O₄SCl) C, H, S, Cl.

(c) N-(4-Fluorophenyl) 4-(carbomethoxy)-2-methylbenzenesulfonamide. Thetitle compound was prepared from 4-fluoroaniline and methyl4-(chlorosulfonyl)-3-methylbenzoate as previously described in Example15(b). ¹H NMR (CDCl₃) δ: 2.68 (s, 3H), 3.93 (s, 3H), 6.62 (s, 1H), 6.95(m, 3H), 7.93 (m, 3H). Anal. (C₁₅H₁₄NO₄SF) C, H, N, S.

(d) N-4-Fluorophenyl) 4-(hydroxymethyl)-2-methylbenzenesulfonamide. Toan ice-cold, stirred solution of N-(4-fluorophenyl)4-(carboxymethyl)-3-methylbenzenesulfonamide (1.00 g, 3.09 mmol) in 25mL THF was added diisobutyl aluminum hydride (7 mL, 1.5 M solution intoluene, 10.5 mol). After 30 minutes at 0° C., another 7 mL ofdiisobutyl aluminum hydride was added. When starting material was gone,the reaction was quenched with saturated Rochelle salt, diluted withEtOAc and stirred vigorously until layers separated. The organic layerwas washed with saturated NaCl solution, dried over MgSO₄, and thesolvents removed under reduced pressure. The residue was purified bychromatography on silica (gradient 20:1 to 5:1 CH₂Cl₂/EtOAc). The titlecompound was isolated as a colorless oil, which solidified on standing(798 mg, 87%). ¹H NMR (CDCl₃) δ: 2.63 (s, 3H), 4.73 (s, 2H), 6.57 (s,1H), 6.94 (m, 4H), 7.24 (d, 1H, J=8.1 Hz), 7.30 (s, 1H), 7.86 (d, 1H,J=8.1 Hz). Anal. (C₁₄H₁₄NO₃SF) C, H, N, S.

(e) N-(4-Fluorophenyl) 4-(bromomethyl)-2-methylbenzenesulfonamide. Thetitle compound was prepared from N-(4-fluorophenyl)4-(hydroxymethyl)-2-methylbenzenesulfonamide in the manner described inExample 1(c). ¹H NMR (CDCl₃) δ: 2.62 (s, 3H), 4.42 (s, 2H), 6.55 (s,1H), 6.89-7.01 (m, 4H), 7.30 (d, 2H, J=8.1 Hz), 7.84 (d, 1H, J=8.1 Hz).Anal. (C₁₄H₁₃NO₂SBrF) C, H, N, S, Br.

(f) N-(4-Fluorophenyl)4-[([5-cyano-6-(5-[2-thienyl]thien-2-yl)-4(3H)-oxopyrimidin-2-yl]thio)methyl]-2-methylbenzenesulfonamide (17). The title compound was preparedfrom 5-cyano-6-[5-(2-thienyl)thien-2-yl]-4(3H)-oxopyrimidine-2-thiol andN-(4-fluorophenyl) 4-(bromomethyl)-2-methylbenzenesulfonamide asdescribed in Example 1(d). ¹H NMR (DMSO-d₆) δ: 2.52 (s, 3H), 4.45 (s,2H), 6.99 (m, 4H), 7.15 (t, 1H, J=4.8 Hz), 7.46 (m, 4H), 7.65 (d, 1H,J=4.8 Hz), 7.76 (d, 1H, J=8.1 Hz), 8.12 (d, 1H, J=3.7 Hz), 10.32 (s,1H). Anal. (C₂₇H₁₉N₄O₃SF.1.1H₂O.0.5EtOAc) C, H, N, S.

Example 18

(a) N-[4-(Carbomethoxy)phenyl] 4-fluorobenzenesulfonamide. To a stirredsolution of methyl 4-aminobenzoate (1.5 g, 9.9 mmol) and pyridine (0.8mL, 9.9 mmol) in CH₂Cl₂ was added 4-fluorobenzenesulfonyl chloride (1.93g, 9.9 mmol). After stirring overnight at RT, the reaction mixture wasdiluted with CH₂Cl₂, and washed sequentially with 0.5N HCl, saturatedNaHCO₃ solution, and saturated NaCl solution, dried over MgSO₄, andconcentrated under reduced pressure. The residue was purified bychromatography on silica (CH₂Cl₂) to give the title compound as a paleyellow solid (2.42 g, 79%). ¹H NMR (CDCl₃) δ: 3.88 (s, 3H), 7.13 (m,5H), 7.84 (dd, 2H, J=9.0, 5.1 Hz), 7.93 (d, 2H, J=8.8 Hz). Anal.(C₁₄H₁₂NO₄SF) C, H, N, S.

(b) N-[4-(Hydroxymethyl)phenyl] 4-fluorobenzenesulfonamide. The titlecompound was prepared by the reduction of N-[4-(carbomethoxy)phenyl](4-fluorophenyl)benzenesulfonamide as generally described in Example17(d). ¹H NMR (CDCl₃) δ: 4.62 (s, 2H), 7.03 (br s, 1H), 7.08 (m, 4H),7.24 (d, 2H, J=8.5 Hz), 7.77 (dd, 2H, J=8.8, 5.1 Hz).

(c)N-(4-[([5-Cyano-6-(5-[2-thienyl]thien-2-yl)-4(3H)-oxopyrimidin-2-yl]thio)methyl]phenyl) 4-fluorobenzenesulfonamide (18). The title compound wasprepared from5-cyano-6-[5-(2-thienyl)thien-2-yl]-4(3H)-oxopyrimidine-2-thiol andN-[4-(bromomethyl)phenyl] 4-fluorobenzenesulfonamide as in Example 1(d).¹H NMR (DMSO-d₆) δ: 4.36 (s, 2H), 7.02 (d, 2H, J=8.5 Hz), 7.14 (t, 1H,J=3.7 Hz), 7.73 (m, 4H), 7.48 (m, 2H), 7.64 (d, 1H, J=4.8 Hz), 7.73 (dd,2H, J=9.2, 5.1 Hz), 8.13 (d, 1H, J=3.7 Hz), 10.32 (s, 1H). Anal.(C₂₆H₂₇N₄O₃S₄F.0.6EtOAc.0.8H₂O) C, H, N, S.

Example 19

(a) Methyl 4-amino-2-chlorobenzoate. A solution of methyl2-chloro-4-nitrobenzoate (4.5 g, 20.9 mmol) and tin (II) chloridedihydrate (18.84 g, 83.5 mmol) in 100 mL EtOAc was heated cautiously to70° C. for 30 minutes. After cooling, the reaction mixture was dilutedwith EtOAc and washed with H₂O (2×), then with saturated NaCl solution,dried over MgSO₄ and concentrated under reduced pressure. In thismanner, the title compound was isolated as an off-while solid (99%). ¹HNMR (CDCl₃) δ: 3.86 (s, 3H), 4.10 (br s, 2H), 6.53 (d, 1H, J=8.5 Hz withfine splitting), 6.70 (s, 1H, with fine splitting), 7.78 (d, 1H, J=8.5Hz). Anal. (C₈H₈NO₂Cl) C, H, N, Cl.

(b) Methyl 4-(chlorosulfonyl)-2-chlorobenzoate. The procedure describedin Chem. Ber. 90 (1957), 841, was used to convert methyl4-amino-2-chlorobenzoate to the title sulfonyl chloride in quantitativeyield. ¹H NMR (CDCl₃) δ: 4.00 (s, 3H), 8.00 (AB, 2H, J=8.1 Hz), 8.13 (s,1H, with fine splitting). Anal. (C₈H₈O₄SCl) C, H, S, Cl.

(c) N-(4-Fluoropbenyl) 4-(carbomethoxy)-3-chlorobenzenesulfonamide. Thetitle compound was prepared from 4-fluoroaniline and methyl4-(chlorosulfonyl)-2-chlorobenzoate in the manner described in Example15(b). ¹H NMR (CDCl₃) δ: 3.95 (s, 3H), 6.63 (s, 1H), 6.95-7.08 (m, 4H),7.58 (d, 1H J=8.5 Hz, with fine splitting), 7.81 (s, 1H, with finesplitting), 7.84 (d, 1H, J=8.1 Hz). Anal. (C₁₄H₁₁NO₄SClF) C, H, N, S,Cl.

(d) N-(4-Fluorophenyl) 4-(hydroxymethyl)-3-chlorobenzenesulfonamide. Thetitle compound was prepared by the reduction of N-(4-fluorophenyl)4-(carbomethoxy)-3-chlorobenzenesulfonamide as described in Example17(d). ¹H NMR (CDCl₃) δ: 4.82 (s, 2H), 6.51 (s, 1H), 6.94-7.07 (m, 4H),7.61 (AB, 2H, J=8.1 Hz, with fine splitting), 7.71 (s, 2H, with finesplitting). Anal. (C₁₃H₁₁NO₃SClF) C, H, N, S, Cl.

(e) N-(4-Fluorophenyl) 4-(bromomethyl)-3-chlorobenzenesulfonamide. Thetitle compound was prepared from N-(4-fluorophenyl)4-(hydroxymethyl)-3-chlorobenzenesulfonamide as described in Example1(c). ¹H NMR (CDCl₃) δ: 4.54 (s, 2H), 6.66 (s, 1H), 6.95-7.09 (m, 4H),7.53 (AB, 2H, J=8.1 Hz, with fine splitting), 7.77 (s, 1H with finesplitting). Anal. (C₁₃H₁₀NO₂SBrCl) C, H, N, S, Br, Cl.

(f) N-(4-Fluorophenyl)4-[([5-cyano-6-(5-[2-thienyl]thien-2-yl)-4(3H)-oxopyrimidin-2-yl]thio)methyl]-3-chlorobenzenesulfonamide(19). The title compound was prepared from5-cyano-6-[5-(2-thienyl)thien-2-yl]-4(3H)-oxopyrimidine-2-thiol andN-(4-fluorophenyl) 4-(bromomethyl)-3-chlorobenzenesulfonamide asdescribed in Example 1(d). ¹H NMR (DMSO-d₆) δ: 4.48 (s, 2H), 7.02 (m,4H), 7.13 (t, 1H, J=3.7 Hz), 7.42 (m, 2H), 7.59 (m, 2H), 7.76 (s, 1H,with fine splitting), 7.80 (d, 1H, J=8.1 Hz), 8.04 (d, 1H, J=4.0 Hz),10.32 (s, 1H). Anal. (C₂₆H₁₆N₄O₃S₄ClF.1.5H₂O) C, H, N,S, Cl.

Example 20

(a) N-[4-(Carbomethoxy)phenyl]-N-methyl 4-fluorobenzenesulfonamide. Astirred solution of N-[4-(carbomethoxy)phenyl]4-fluorobenzenesulfonamide (440 mg, 1.42 mmol), methyl iodide (0.132 mL,2.13 mmol), and N,N-diisopropylethylamine (0.372 mL, 2.13 mmol) inCH₂Cl₂ was heated at reflux for 18 hours. Another 0.10 mL of methyliodide and 0.2 mL of N,N-diisopropylethylamine were added, and heatingcontinued for 4 hours more. After cooling, the mixture was diluted withEtOAc and washed sequentially with 0.5N HCl, then saturated NaClsolution. The organic layer was separated, dried over MgSO₄ and thesolvent removed under reduced pressure. The residue was purified bychromatography on silica (CH₂Cl₂) to give the title compound (386 mg,84%) as a white solid. ¹H NMR (CDCl₃) δ: 3.20 (s, 3H), 3.95 (s, 3H),7.12 (dd, 2H, J=8.8, 8.5 Hz), 7.21 (d, 2H, J=8.8 Hz), 7.53 (dd, 2H,J=8.8, 5.1 Hz), 7.98 (d, 2H, J=8.8 Hz). Anal. (C₁₅H₁₄NO₄SF) C, H, N, S.

(b) N-[4-(Hydroxymethyl)phenyl]-N-methyl 4-fluorobenzenesulfonamide. Thetitle compound was prepared by the reduction ofN-[4-(carbomethoxy)phenyl]-N-methyl 4-fluorobenzenesulfonamide asdescribed in Example 17(d). ¹H NMR (CDCl₃) δ: 3.17 (s, 3H), 4.70 (s,2H), 7.09 (d, 2H, J=8.5 Hz), 7.15 (d, 2H, J=8.5 Hz), 7.32 (d, 2H, J=8.5Hz), 7.56 (dd, 2H, J=9.1, 5.1 Hz).

(c) N-[4-(Bromomethyl)phenyl]-N-methyl 4-fluorobenzenesulfonamide. Thetitle compound was prepared from N-[4-(hydroxymethyl)phenyl]-N-methyl4-fluorobenzenesulfonamide as described in Example 1(c). ¹H NMR (CDCl₃)δ: 3.16 (s, 3H), 4.47 (s, 2H), 7.08 (d, 2H, J=8.5 Hz), 7.14 (dd, 2H,J=8.8, 8.5 Hz), 7.31 (d, 2H, J=8.5 Hz), 7.55 (dd, 2H, J=8.8, 5.1 Hz).Anal. (C₁₄H₁₃NO₂SBrF) C, H, N, S, Br.

(d) N-(4-[([5Cyano-6(5-[2-thienyl]thien-2-yl)-4(3H)-oxopyrimidin-2-yl]thio)methyl]phenyl)-N-methyl4-fluorobenzenesulfonamide (20). The title compound was prepared from5-cyano-6-[5-(2-thienyl)thien-2-yl]-4(3H)-oxopyrimidine-2-thiol andN-[4-(bromomethyl)phenyl]-N-methyl 4-fluorobenzenesulfonamide asdescribed in Example 1(d). ¹H NMR (DMSO-d₆) δ: 3.08 (s, 3H), 4.53 (s,2H), 7.07 (d, 2H, J=8.5 Hz), 7.15 (t, 1H, J=3.7 Hz), 7.35 (dd, 2H,J=8.8, 8.8 Hz), 7.45 (d, 2H, J=8.5 Hz), 7.53 (m, 4H), 7.66 (d, 1H, J=4.0Hz), 8.19 (d, 1H, J=4.1 Hz). Anal. (C₂₇H₁₉N₄O₃S₄F.0.5H₂O) C, H, N, S.

Example 21

(a) N-[4-(Carbomethoxy)-3-chlorophenyl] 4-fluorobenzenesulfonamide. Thetitle compound was prepared from methyl 4-amino-2-chlorobenzoate and4-fluorobenzenesulfonyl chloride as previously described in Example15(b). ¹H NMR (CDCl₃) δ: 3.89 (s, 3H), 7.05 (dd, 1H, J=8.5, 2.2 Hz),7.14-7.20 (m, 3H), 7.78 (d, 1H, J=8.5 Hz), 7.87 (dd, 2H, J=7.0, 5.0 Hz).Anal. (C₁₄H₁₁NO₄SClF) C, H, N, S, Cl.

(b) N-[3-Chloro-4-(hydroxymethyl)phenyl] 4-fluorobenzenesulfonamide. Thetitle compound was prepared by the reduction ofN-[4-(carbomethoxy)-3-chlorophenyl] 4-fluorobenzenesulfonamide asdescribed in Example 17(c). ¹H NMR (CDCl₃) δ: 4.70 (s, 2H), 6.97 (d, 1H,J=8.5 Hz), 7.05 (s, 1H), 7.14 (m, 3H), 7.35 (d, 1H, J=8.5 Hz), 7.81 (dd,2H, J=8.8, 5.1 Hz). Anal. (C₁₃H₁₁NO₃SClF) C, H, N, S, Cl.

(c)N-(3-Chloro-4-[([5-cyano-6-(5-[2-thienyl]thien-2-yl)-4(3H)-oxopyrimidin-2-yl]thio)methyl]phenyl)4-fluorobenzenesulfonamide (21). The title compound was prepared from5-cyano-6-[5-(2-thienyl)thien-2-yl]-4(3H)-oxopyrimidine-2-thiol andN-[4-(bromomethyl)-3-chlorophenyl] 4-fluorobenzenesulfonamide asdescribed in Example 1(d). ¹H NMR (DMSO-d₆) δ: 4.42 (s, 2H), 7.01 (d,1H, J=8.1 Hz), 7.16 (m, 2H), 7.33 (dd, 2H, J=8.8, 8.8 Hz), 7.49 (m, 3H),7.64 (d, 1H, J=4.8 Hz), 7.80 (dd, 2H, J=8:9, 5.1 Hz), 8.10 (d, 1H, J=4.1Hz), 10.62 (s, 1H). Anal. (C₂₆H₁₆N₄O₃S₄ClF.0.5H₂O) C, H, N, S.

Example 22

(a) N-[4-(Carbomethoxy)-2-methylphenyl] 4-fluorobenzenesulfonamide. Thetitle compound was prepared from methyl 4-amino-3-methylbenzoate and4-fluorobenzenesulfonyl chloride in the manner earlier described inExample 15(b). ¹H NMR (CDCl₃) δ: 2.08 (s, 3H), 3.92 (s, 3H), 6.61 (s,1H), 7.13 (dd, 2H, J=8.5, 8.5 Hz), 7.46 (d, 1H, J=8.1 Hz), 7.81 (m, 4H).Anal. (C₁₅H₁₄NO₄SF) C, H, N, S.

(b) N-[4-(Hydroxymethyl)-2-methylphenyl] 4-fluorobenzenesulfonamide. Thetitle compound was prepared by the reduction ofN-[4(carbomethoxy)-2-methylphenyl] 4-fluorobenzenesulfonamide asdescribed above in Example 17(d). ¹H NMR (CDCl₃) δ: 2.01 (s, 3H), 4.62(s, 2H), 6.37 (s, 1H), 7.13 (m, 3H), 7.25 (dd, 2H, J=8.1, 5.8 Hz), 7.73(dd, 2H, J=8.8, 5.1 Hz). Anal. (C₁₄H₁₄NO₃SF.0.1H₂O) C, H, N, S.

(c)N-(4-[([5-Cyano-6-(5-[2-thienyl]thien-2-yl)4(3H)-oxopyrimidin-2-yl]thio)methyl]-2-methylphenyl)4-fluorobenzenesulfonamide (22). The title compound was prepared from5-cyano-6-[5-(2-thienyl)thien-2-yl]-4(3H)-oxopyrimidine-2-thiol andN-[4-(bromomethyl)-2-methylphenyl] 4-fluorobenzenesulfonamide asdescribed in Example 1(d). ¹H NMR (DMSO-d₆) δ: 1.95 (s, 3H), 4.44 (s,2H), 6.89 (d, 1H, J=8.1 Hz), 7.19 (m, 3H), 7.31 (s, 1H), 7.35 (d, 2H,J=8.8 Hz), 7.53 (d, 2H, J=3.7 Hz), 7.67 (dd, 2H, J=8.8, 5.1 Hz), 8.19(d, 1H, J=4.1 Hz), 9.63 (s, 1H). Anal. (C₂₇H₁₉N₄O₃S₄F.0.5H₂O) C, H, N,S.

Example 23

(a) N-[4-(Carbomethoxy)-3-methylphenyl] 4-fluorobenzenesulfonamide. Thetitle compound was prepared from methyl 4-amino-2-methylbenzoate and4-fluorobenzenesulfonyl chloride as described in Example 15(b). ¹H NMR(CDCl₃) δ: 3.84 (s, 3H), 3.85 (s, 3H), 6.57 (d, 1H, J=8.5 Hz with finesplitting), 6.82 (s, 1H, with fine splitting), 6.91 (s, 1H), 7.14 (dd,2H, J=8.8, 8.5 Hz), 7.71 (d, 1H, J=8.5 Hz), 7.84 (dd, 2H, J=8.8, 4.8Hz). Anal. (C₁₅H₁₄NO₅SF.0.2H₂O) C, H, N, S.

(b) N-[4-(Hydroxymethyl)-3-methoxyphenyl] 4-fluorobenzenesulfonamide.The title compound was prepared by the reduction ofN-[4-(carbomethoxy)-3-methoxyphenyl] 4-fluorobenzenesulfonamide aspreviously described in Example 17(d). ¹H NMR (CDCl₃) δ: 3.82 (s, 3H),4.60 (s, 2H), 6.48 (d, 1H, J=8.1 Hz with fine splitting), 6.59 (s, 1H),6.77 (s, 1H with fine splitting), 7.09-7.28 (m. 3H), 7.77 (m, 2H). Anal.(C₁₄H₁₄NO₄SF.0.15PhCH₃) C, H, N, S.

(c)N-(4-[([5-Cyano-6-(5-[2-thienyl]thien-2-yl)-4(3H)-oxopyrimidin-2-yl]thio)methyl]-3-methoxyphenyl)4-fluorobenzenesulfonamide (23). The title compound was prepared from5-cyano-6-[5-(2-thienyl)thien-2-yl]-4(3H)-oxopyrimidine-2-thiol andN-[4-(bromomethyl)-3-methoxyphenyl] 4-fluorobenzenesulfonamide asdescribed in Example 1(d). ¹H NMR (DMSO-d₆) δ: 3.75 (s, 3H), 4.37 (s,3H), 4.37 (s, 3H), 6.60 (d, 1H, J=8.1 Hz with fine splitting), 6.79 (s,1H, with fine splitting), 7.15 (dd, 1H, J=4.8, 3.7 Hz), 7.28 (d, 1H,J=8.1 Hz), 7.53 (m, 2H), 7.67 (d, 1H, J=5.2 Hz), 7.82 (dd, 2H, J=8.8,5.1 Hz), 8.19 (d, 1H, J=4.8 Hz), 10.40 (s, 1H). Anal.(C₂₇H₁₉N₄O₄S₄F.0.8MeOH) C, H, N, S.

Example 24

(a) Methyl 4-amino-3-methylbenzoate. A stirred solution of4-amino-3-methoxybenzoic acid (2.50 g, 14.97 mmol) in MeOH saturatedwith HCl gas was heated at reflux overnight. The volatiles were removedunder reduced pressure and the residue dissolved in EtOAc, washed withsaturated NaHCO₃ solution, dried over MgSO₄ and concentrated underreduced pressure. The title compound was isolated as a tan solid (97%).¹H NMR (CDCl₁) δ: 3.87 (s, 3H), 3.91 (s, 3H), 4.5 (br s, 2H), 6.73 (d,1H, J=8.1 Hz), 7.46 (s, 1H, with fine splitting). Anal. (C₁₂H₁₁NO₃) C,H, N.

(b) N-[4-(Carbomethoxy)-2-methoxyphenyl] 4-fluorobenzenesulfonamide. Thetitle compound was prepared from methyl 4-amino-3-methylbenzoate and4-fluorobenzenesulfonyl chloride as described in Example 15(b). ¹H NMR(CDCl₃) δ: 3.79 (s, 3H), 3.88 (s, 3H), 7.1 (m, 2H), 7.28 (s, 1H), 7.43(s, 1H with fine splitting), 7.60 (m, 2H), 7.83 (m, 2H). Anal.(C₁₅H₁₄NO₅SF) C, H, N, S.

(c) N-[4-(Hydroxymethyl)-2-methoxyphenyl] 4-fluorobenzenesulfonamide.The title compound was prepared by the reduction ofN-[4-(carbomethoxy)-2-methoxyphenyl] 4-fluorobenzenesulfonamide asdescribed in Example 17(d). ¹H NMR (DMSO-d₆) δ: 3.51 (s, 3H), 4.66 (d,2H, J=5.9 Hz), 5.22 (t, 1H, J=5.9 Hz), 6.87 (d, 1H, J=8.1 Hz), 6.89 (s,1H), 7.19 (d, 1H, J=8.1 Hz), 7.41 (dd, 2H, J=8.8, 8.8 Hz), 7.78 (dd, 2H,J=8.8, 5.1 Hz), 9.53 (s, 1H). Anal. (C₁₄H₁₄NO₄SF) C, H, N, S.

(d)N4-[([5-Cyano-6-(5-[2-thienyl]thien-2-yl)-4(3H)-oxopyrimidin-2-yl]thio)methyl]-2-methoxyphenyl)4-fluorobenzenesulfonamide (24). The title compound was prepared from5-cyano-6-[5-(2-thienyl)thien-2-yl]-4(3H)-oxopyrimidine-2-thiol andN-[4-(bromomethyl)-2-methoxyphenyl] 4-fluorobenzenesulfonamide asdescribed in Example 1(d). ¹H NMR (DMSO-d₆) δ: 3.39 (s, 3H), 4.43 (s,2H), 6.98 (d, 1H, J=8.1 Hz), 7.07 (s, 1H), 7.14 (d 2H, J=8.7 Hz), 7.31(dd, 2H, J=8.7, 8.1 Hz), 7.51 (m, 2H), 7.68 (m, 3H), 8.17 (d, 1H, J=3.7Hz), 9.56 (s, 1H). Anal. (C₂₇H₁₉N₄O₄S₄F.0.2H₂O.0.4EtOAc) C, H, N, S.

Example 25

(a) N-[3-(Carbomethoxy)phenyl] 4-fluorobenzenesulfonamide. The titlecompound was prepared from methyl 3-aminobenzoate and4-fluorobenzenesulfonyl chloride as described in Example 15(b). ¹H NMR(CDCl₃) δ: 1.38 (t, 3H, J=7.0 Hz), 4.37 (q, 2H, J=7.0 Hz), 6.89 (s, 1H),7.12 (t, 2H, J=8.5 Hz), 7.38 (m, 2H), 7.67 (s, 1H, with fine splitting),7.80 (m, 3H). Anal. (C₁₄H₁₂NO₄SF) C, H, N, S.

(b) N-[3-(Hydroxymethyl)phenyl] 4-fluorobenzenesulfonamide. The titlecompound was prepared by the reduction of N-[3-(carbomethoxy)phenyl](4-fluorophenyl) benzenesulfonamide as described in Example 17(d). ¹HNMR(CDCl₃) δ: 4.39 (s,2H), 6.75 (s, 1H), 6.98 (d, 1H, J=8.1 Hz),7.09-7.65 (m, 5H), 7.78 (dd, 2H, J=8.8, 4.8 Hz).

(c)N(3-[([5-Cyano-6-(5-[2-thienyl]thien-2-yl)-4(3H)-oxopyrimidin-2-yl]thio)methyl]phenyl)4-fluorobenzenesulfonamide. The title compound was prepared from5-cyano-6-[5-(2-thienyl)thien-2-yl]-4(3H)-oxopyrimidine-2-thiol andN-[3-(bromomethyl)phenyl] 4-fluorobenzenesulfonamide as described inExample 1(d). ¹H NMR (DMSO-d₆) δ: 4.51 (s, 2H), 7.01 (d, 1H, J=7.7 Hz),7.20-7.35 (m, 6H), 7.57 (d, 1H, J=3.7 Hz), 7.59 (s, 1H), 7.72 (d, 1H,J=4.8 Hz), 7.78 (dd, 2H, J=8.8, 5.1 Hz), 8.24 (d, 1H, J=4.4 Hz), 10.42(s 1H). Anal. (C₂₆H₁₇N₄O₃S₄F0.5 CH₂Cl₂) C, H, N, S.

Example 26

(a) Methyl 3-(iodopbenyl)-3-oxopropionate. The title compound wasprepared from 3′-iodoacetophenone in a manner described in Example 4(c).¹H NMR (CDCl₃) δ: 8.27 (1H, dd, J=1.5, 1.8 Hz), 7.95-7.87 (2H, m), 7.22(1H, dd, J=1.5, 7.7 Hz), 3.97 (2H, s), 3.76 (3H, s). Anal. (C₁₀H₉O₃I) C,H, I.

(b) 6(3-Iodophenyl)-4(3H)-oxopyrimidine-2-thiol. The title compound wasprepared from methyl 3-(iodophenyl)-3-oxopropionate and thiourea aspreviously described in Example 4(d). ¹H NMR (DMSO-d₆) δ: 12.60 (1H, brs), 12.57 (1H, br s), 8.11 (1H, s), 7.96 (1H, d, J=8.1 Hz), 7.75 (1H, d,J=8.1 Hz),7.33 (1H, t, J=8.1 Hz), 6.17 (1H, s). Anal. (C₁₀H₇N₂OSI) C, H,N, S, I.

(c) Methyl5-[5-([6-(3-iodophenyl)-4(3H)-oxopyrimidin-2-yl]thio)pentyl]thiophene-2-carboxylate.The title compound was prepared from6-(3-iodophenyl)-4(3H)-oxopyrimidine-2-thiol and methyl5-(5-bromopentyl)thiophene-2-carboxylate as previously described inExample 1(d). ¹H NMR (DMSO-d₆) δ: 12.75 (1H, br), 8.40 (1H, s), 8.04(1H, d, J=8.1 Hz), 7.83 (1H, d, J=8.1 Hz), 7.59 (1H, d, J=3.7 Hz), 7.25(1H, t, J=8.1 Hz), 6.92 (1H, d, J=3.7 Hz), 6.70 (1H, s), 3.77 H, s),3.19 (2H, t, J=7.3 Hz), 2.85 (2H, t, J=7.4 Hz), 1.79-1.64 (4H, m),1.52-1.42 (2H, m). Anal. (C₂₁H₂₁N₂O₃S₂I) C, H, N, S, I.

(d) Methyl5-[5-([6-(3-cyanophenyl)-4(3H)-oxopyrimidin-2-yl]thio)pentyl]thiophene-2-carboxylate.To a solution of methyl5-[5-([6-(3-iodophenyl)-4(3H)-oxopyrimidin-2-yl]thio)pentyl]thiophene-2-carboxylate(505 mg, 1 mmol) in THF (25 mL) were addedtetrakis(triphenylphosphine)palladium (188 mg, 0.2 mmol) and KCN (195mg, 3 mmol). The resulting mixture was heated overnight at reflux underan argon atmosphere. After cooling to room temperature, the precipitatewas removed by filtration and washed with THF (3×10 mL). The combinedfiltrates were subsequently concentrated in vacuo to give a yellow gum,which was purified by flash chromatography. Elution with CH₂Cl₂:CH₃OH(97:3) provided the product as a white solid (378 mg, 92% yield). ¹H NMR(DMSO-d₆) δ: 12.87 (1H, br s), 8.46 (1H, s), 8.35 (1H, d, J=8.1 Hz),7.94 (1H, d, J=7.7 Hz), 7.66 (1H, dd, J=7.7, 8.1 Hz), 7.59 (1H, d, J=3.7Hz), 6.91 (1H, d, J=3.7 Hz), 6.82 (1H, s), 3.77 (3H, s), 3.24 (2H, t,J=7.2 Hz), 2.83 (2H, t, J=7.3 Hz), 1.80-1.63 (4H, m), 1.51-1.41 (2H, m).Anal. (C₂₂H₂₁N₃O₃S₂) C, H, N, S.

(e) Methyl5-[5-([6-(3-[1(H)-tetrazol-5-yl]phenyl)-4(3H)-oxopyrimidin-2-yl]thio)pentyl]thiophene-2-carboxylate. To a solution of methyl5-[5-([6-(3-cyanophenyl)-4(3H)-oxopyrimidin-2-yl]thio)pentyl]thiophene-2-carboxylate(303 mg, 0.7 mmol) in DMF (15 ml) were added sodium azide (672 mg, 10mmol) and ammonium chloride (553 mg, 10 mmol). The reaction mixture washeated at 85° C. for about 16 hours, then poured into water (100 mL).The precipitate that formed was collected by filtration and washed withwater (3×15 mL) to provide the product as an orange solid (252 mg, 76%yield). ¹H NMR (DMSO-d₆) δ: 12.84 (1H, br), 8.73 (1H, s), 8.21 (1H, d,J=8.1 Hz), 8.12 (1H, d, J=7.7 Hz), 7.68 (1H, dd, J=7.7, 8.1 Hz), 7.56(1H, d, J=3.7 Hz), 6.87 (1H, d, J=3.7 Hz), 6.77 (1H, s), 3.76 (3H, s),3.27 (2H, t, J=7.4 Hz), 2.81 (2H, t, J=7.3 Hz), 1.79-1.61 (4H, m),1.53-1.46 (2H, m). Anal. (C₂₂H₂₂N₆O₃S₂.0.3H₂O) C, H, N, S.

(f)5-[5-([6-(3-[1(H)-Tetrazol-5-yl]phenyl)-4(3H)-oxopyrimidin-2-yl]thio)pentyl]thiophene-2-carboxylicacid (26). Compound 26 was prepared by hydrolysis of methyl 5-[5-([6-(3-[1(H)-tetrazol-5-yl]phenyl)-4(3H)-oxopyrimidin-2-yl]thio)pentyl]thiophene-2-carboxylatein analogy to Example 1(e). ¹H NMR (DMSO-d₆) δ: 12.81 (2H, br), 8.74(1H, s), 8.23 (1H, d, J=8.1 Hz), 8.13 (1H, d, J=7.7 Hz), 7.69 (1H, ddJ=7.7, 8.1 Hz), 7.50 (1H, d, J=3.7 Hz), 6.84 (1H, d, J=3.7 Hz), 6.77(1H, s), 3.27 (2H, t, J=7.3 Hz), 2.79 (2H, t, J=7.4 Hz), 1.80-1.61 (4H,m), 1.54-1.46 (2H, m). Anal. (C₂₁H₂₀N₆O₃S₂.1.1H₂O) C, H, N, S.

Example 27

N-(2-[1(H)-indol-3-yl]-ethyl)4-[([5-cyano-6-(5-[2-thienyl]thien-2-yl)-4(3H)-oxopyrimidin-2-yl]thio)methyl]benzenesulfonamide (14). The title compound was prepared from5-cyano-6-[5-(2-thienyl)thien-2-yl]-4(3H)-oxopyrimidine-2-thiol andN-(2-[1(H)-indol-3-yl]-ethyl) 4-(bromomethyl) benzenesulfonamidegenerally as described in Example 1(d). N-(2-[1(H)-indol-3-yl]-ethyl)4-(bromomethyl) benzenesulfonamide was prepared from tryptamine and4-(bromomethyl)benzenesulfonyl chloride as described in Example 15(b).

BIOCHEMICAL ASSAYS

AICARFT Activity

Enzymatic activity was measured spectrophotometrically using a modifiedassay developed by Black et al. (“A Rapid Assay for5-Amino-4-imidazolecarboxamide Ribotide Transformylase,” Anal. Biochiem.90 (1978), 397-401). The reaction volume was 1 mL and contained 50 mMTris-HCl pH 7.5, 25 mM KCl, 20 mM 2-mercaptoethanol, variableconcentrations of test compound in 1% DMSO, 50 mM AICAR(5-aminoimidazole-4-carboxamideribonucleotide), 50 mM N¹⁰-formyltetrathydrofolate (FTHF), and 0.015 mM enzyme. The reaction wasenzyme-initiated and followed by monitoring the increase in absorbanceat 298 nm at 25° C. (ξ₂₉₈=19.7×10³ cm⁻¹M⁻¹). AICARFT inhibitionconstants (K_(i)) were determined from the dependence of thesteady-state catalytic rate upon compound and substrate concentration.The error associated with this assay is less than 10% of a given K_(i)value and is often less than 5%. values for compounds of the inventionprepared as described above are tabulated below.

AICARFT COMPOUND K_(i) (μM) 1 3.5 2 7.2 3 0.28 4 54.7 5 0.27 6 0.43 70.22 8 0.04 9 0.46 10 0.004 11 6.8 12 8.9 13 5.3 14 0.59 15 2.1 16 0.4317 1 18 0.38 19 0.4 20 1.8 21 0.029 22 1 23 0.77 24 1.3 25 1.4 26 0.3727 0.083

While the invention has been illustrated above in terms of preferredembodiments and specific examples, those skilled in the art willrecognize that various changes and modifications can be made withoutdeparting from the spirit and scope of the invention. Thus, theinvention should be understood as not being limited by the foregoingdescription, but as being defined by the appended claims and theirequivalents.

What is claimed is:
 1. A compound having the structure:

wherein: R¹is H or CN; R² is phenyl or thienyl, each of which may be optionally substituted with phenyl, phenoxy, thienyl, tetrazolyl, or 4-morpholinyl; and R³ is phenyl substituted by —SO₂NR⁵R⁶ or —NR⁵SO₂R⁶ and optionally further substituted with lower alkyl, lower alkoxy, or halogen, wherein R⁵ is H or lower alkyl, R⁶ is lower alkyl, heteroarylalkyl, substituted or unsubstituted with one or more substituents chosen from the group consisting of hydroxy, halogens, oxo, alkyl, acyl, sulfonyl, mercapto, alkylthio, alkoxy, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, carboxy, amino, alkylamino, dialkylamino, carbamoyl, aryloxy, heteroaryloxy, arylthio, and heteroarylthio, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, or R³ is

 wherein n is an integer of from 1 to 4, R4 is OH, lower alkoxy, or glutamic-acid or glutamate-alkylester linked through the amine functional group, or a pharmaceutically acceptable salt thereof.
 2. A compound according to claim 1, wherein aryl and heteroaryl are substituted with lower alkyl, alkoxy or halogen.
 3. A compound according to claim 1, wherein aryl is phenyl and heteroaryl is isoquinolyl or indolyl.
 4. A compound selected from the group consisting of: 